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66,641
Dual Luciferase Assay 
1
null
https://www.protocols.io/view/dual-luciferase-assay-cdbrs2m6
vahedilaboratory
TITLE: Dual Luciferase Assay  AUTHORS: vahedilaboratory [DESCRIPTION] Dual luciferase assay WIP [STEPS] 3. Cell Lysing and Luciferase Assay Remove media from each well and wash with PBS; remove the PBS Add 50uL trypsin to each well and coat every cell Leave in incubator for 3 minutes Pipette all cells off of plate...
["Cell Lysing and Luciferase Assay\nRemove media from each well and wash with PBS; remove the PBS\nAdd 50uL trypsin to each well and coat every cell\nLeave in incubator for 3 minutes\nPipette all cells off of plate and add cells to 1.5mL tubes \nSpin down cells at 1,500RPM 4C for 5 minutes to pellet cells \nRemove supe...
null
null
null
dx.doi.org/10.17504/protocols.io.cn2vgd
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
34,192
LyGo cloning
1
dx.doi.org/10.17504/protocols.io.bdmqi45w
https://www.protocols.io/view/lygo-cloning-bdmqi45w
Kristoffer Bach Falkenberg, Cristina Hernandez Rollan, Maja Rennig, Andreas Birk Bertelsen, Morten Norholm
TITLE: LyGo cloning AUTHORS: Kristoffer Bach Falkenberg, Cristina Hernandez Rollan, Maja Rennig, Andreas Birk Bertelsen, Morten Norholm [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Lytic polysaccharide monooxygenses (LPMOs) are enzymes that play a critical role in breaking the chemical bonds of t...
["[Mix reactions]\nPrepare a reaction mix in the following proportions: AB1ComponentVolume (μL)2LyGo vector13LyGo fragment34FastDigest SapI0.5510x T4 ligase1610x FastDigest buffer1710x T4 buffer18MQ2.59Total10\nAB1ComponentVolume (μL)2LyGo vector13LyGo fragment34FastDigest SapI0.5510x T4 ligase1610x FastDigest buffer1...
100,280
Integrated laser ablation and droplet collection system for efficient single cell isolation
0
dx.doi.org/10.17504/protocols.io.kxygxy3ewl8j/v1
https://www.protocols.io/view/integrated-laser-ablation-and-droplet-collection-s-dd6y29fw
Liang Chen, Andrey V. Liyu, Yumi Kwon, Ying Zhu, Ljiljana.PasaTolic
TITLE: Integrated laser ablation and droplet collection system for efficient single cell isolation AUTHORS: Liang Chen, Andrey V. Liyu, Yumi Kwon, Ying Zhu, Ljiljana.PasaTolic [DESCRIPTION] State-of-the-art omics and imaging technologies for transcriptome, epigenome, and proteome measurements at the single-cell level ...
["[Laser capture microdissection (LCM)] Install in-house built robotic arm, which is used in the generation and delivery of a captured droplet.", "[Laser capture microdissection (LCM)] Wash the Syringe and glass capillary with MilliQ water", "[Laser capture microdissection (LCM)] Turn on the PALM MicroBeam system", "[L...
109,018
Nested PCR amplification of Salmonella Typhi from extracted wastewater concentrate
0
dx.doi.org/10.17504/protocols.io.36wgqnw3ogk5/v1
https://www.protocols.io/view/nested-pcr-amplification-of-salmonella-typhi-from-dnp25dqe
Shannon Fitz, Alex Shaw, michael Owusu, Dilip Abraham, Anton Spadar
TITLE: Nested PCR amplification of Salmonella Typhi from extracted wastewater concentrate AUTHORS: Shannon Fitz, Alex Shaw, michael Owusu, Dilip Abraham, Anton Spadar [DESCRIPTION] This standard operating procedure describes nested PCR protocols for the generation of amplicons from V. cholerae and S. typhi.    PCR pr...
["[Primer preparation step] Workstation Preparation\n\nClean the PCR areas before setting up the reactions; clean room for the master mix and an area for the addition of the template. \nClean the working area and pipettors with an approved DNAse inhibitor solution.\nClean the Class II biosafety cabinet and pipettes wit...
null
null
null
dx.doi.org/10.17504/protocols.io.scpeavn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol outlines our preparation of single-cell suspension from surgically acquired fresh human adult brain tissue. This method is adapted from Habib et al<sup>1</sup>.</p> <p> </p> <p>1          Habib, N.<em> et al.</em> Massively parallel single-nucleus RNA-seq with D...
[]
67,387
Bewertungen von Diaetoxil-Kapseln - Effektive Inhaltsstoffe wirken beim Abnehmen!
1
dx.doi.org/10.17504/protocols.io.n2bvj645wlk5/v1
https://www.protocols.io/view/bewertungen-von-diaetoxil-kapseln-effektive-inhalt-cd23s8gn
natedif
TITLE: Bewertungen von Diaetoxil-Kapseln - Effektive Inhaltsstoffe wirken beim Abnehmen! AUTHORS: natedif [DESCRIPTION] ➢ Produktname – Diaetoxil ➢ Nebenwirkungen – NA ➢ Zusammensetzung — Natürliche organische Verbindung ➢ Vorteile – Hilft, Gewicht zu reduzieren und die Energie zu steigern! ➢ Verfügbarkeit — Auf Lag...
[]
20,015
Preparation of the adenylated oligonucleotide for mapping 3′OH RNA termini
null
dx.doi.org/10.17504/protocols.io.xspfndn
null
Matus Valach
TITLE: Preparation of the adenylated oligonucleotide for mapping 3′OH RNA termini AUTHORS: Matus Valach [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Adapted from the protocol for oligonucleotide 5′ adenylation (E2610) of </span><span style = "font-style:italic;">New England Biolabs</span><s...
["Mix the following components (20 μL): ABC1ComponentAmount [μL]Final concentration25′ phosphorylated DNA oligo [50 μM]410 μM310× 5′ DNA Adenylation buffer21×4ATP [1 mM]20.1 mM5Mth RNA ligase [50 pmol/µL]410 µM6RNase-free water (ddH2O)8\nABC1ComponentAmount [μL]Final concentration25′ phosphorylated DNA oligo [50 μM]41...
27,501
Effective Lonza 4D Nucleofection with Inexpensive Homemade Buffers
null
dx.doi.org/10.17504/protocols.io.64mhgu6
https://www.protocols.io/view/effective-lonza-4d-nucleofection-with-inexpensive-64mhgu6
Eric Danner
TITLE: Effective Lonza 4D Nucleofection with Inexpensive Homemade Buffers AUTHORS: Eric Danner [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol uses the Lonza 4D nucleofection system. Here I present protocols that reduce the price dramatically of using Lonza 4D equipment. </div><div cl...
["[Prepare Reagents]\nMake Nucleofection Buffer (AmaxaV): Stocks can be stored RT for a long time (we use ours longer than 6 months). Making working solutoin from stock solution, vacuum filter through 0.45um filter. This should be stored at 4C. Discard after 3 months or when you see white precipitate. ABCDEFG1Stoc...
21,662
Analysis of the Derivatives of Sialic Acid in Pig Tissues
null
dx.doi.org/10.17504/protocols.io.zd6f29e
null
Chon-Ho Yen
TITLE: Analysis of the Derivatives of Sialic Acid in Pig Tissues AUTHORS: Chon-Ho Yen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Sialic acid is a generic term for the N- or O-substituted derivatives of neuraminic acid, a monosaccharide with a nine-carbon backbone. N-glycolylneuraminic acid (Ne...
[]
51,966
A Difference-in-Differences Analysis of New Persistent Opioid Use After Surgery
1
dx.doi.org/10.17504/protocols.io.bwy6pfze
https://www.protocols.io/view/a-difference-in-differences-analysis-of-new-persis-bwy6pfze
Ryan Howard, Craig Brown, Vidhya Gunaseelan, Michael Englesbe, Jennifer Waljee, Mark Bicket, Andrew Ryan, Chad Brummett
TITLE: A Difference-in-Differences Analysis of New Persistent Opioid Use After Surgery AUTHORS: Ryan Howard, Craig Brown, Vidhya Gunaseelan, Michael Englesbe, Jennifer Waljee, Mark Bicket, Andrew Ryan, Chad Brummett [DESCRIPTION] The Opioid Prescribing Engagement Network (OPEN) at the University of Michigan was establ...
["[Brief Rationale and Hypothesis] Brief Summary: The Opioid Prescribing Engagement Network (OPEN) at the University of Michigan was established in 2016 with the goal of reducing excessive opioid prescribing after surgery using evidence-based prescribing guidelines. Beginning in July 2016, OPEN began a statewide qualit...
80,893
Methodology for Inputs-Oriented VRS DEA in dairy farms
1
dx.doi.org/10.17504/protocols.io.ewov1o6z7lr2/v1
https://www.protocols.io/view/methodology-for-inputs-oriented-vrs-dea-in-dairy-f-cs85why6
C A Zuniga-Gonzalez, Jaramillo
TITLE: Methodology for Inputs-Oriented VRS DEA in dairy farms AUTHORS: C A Zuniga-Gonzalez, Jaramillo [DESCRIPTION] The methodology protocol for Inputs-Oriented VRS DEA in dairy farms describes the steps and procedures for data collection with the verbal consent of the producers according to the declaration included i...
["[Protocol for the collection and processing of data with the verbal consent of the producers] The data collection procedure was as follows: a) identification of the regions of the republic with the highest volume of milk production. A questionnaire was designed to collect information from the selected producers. Dur...
64,710
Illumina double-stranded DNA dual indexing for ancient DNA
1
dx.doi.org/10.17504/protocols.io.4r3l287x3l1y/v3
https://www.protocols.io/view/illumina-double-stranded-dna-dual-indexing-for-anc-cbfesjje
Raphaela Stahl, Christina Warinner, Irina Velsko, Eleftheria Orfanou, Franziska Aron, Guido Brandt
TITLE: Illumina double-stranded DNA dual indexing for ancient DNA AUTHORS: Raphaela Stahl, Christina Warinner, Irina Velsko, Eleftheria Orfanou, Franziska Aron, Guido Brandt [DESCRIPTION] This protocol converts partially completed double-stranded DNA libraries e.g. from: Non-UDG treated double-stranded ancient DNA li...
["[Indexing preparation (aDNA library preparation room)] Calculate the total number of DNA molecules (total copy number) DNA concentration in each library based on qPCR performed at the end of library preparation (see Before Start). Do not use more than 1.5x10^10 copies per indexing reaction. Adjust the amount of DNA u...
41,758
C-19 protocol
4
null
https://www.protocols.io/view/c-19-protocol-bkz6kx9e
yalhiyari
TITLE: C-19 protocol AUTHORS: yalhiyari [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">A rapid covid screening taking 30sec- 1 min once placed in DOCI machine. Also does not require any biological sample manipulation.</div></div> [STEPS] ?. Centrifuge: 3500 33, 5 min, 22 10 ?. Need pipet tips an...
["Centrifuge: 3500 33, 5 min, 22 10", "Need pipet tips and 20-200ul pipet, 96well glass bottom plate Fishersci 07-000-633\n[Aliquot 50ul of supernatant 1 well per sample into 96 well glass bottom plate ]", "Stick 96 well plate into machine for viral detection. 30 sec-1 min per run"]
44,764
3.5 Applications of G-Trap Assay
3
dx.doi.org/10.17504/protocols.io.bpx4mpqw
https://www.protocols.io/view/3-5-applications-of-g-trap-assay-bpx4mpqw
Peter Simons, Virginie Bondu, Angela Wandinger-Ness, Tione Buranda
TITLE: 3.5 Applications of G-Trap Assay AUTHORS: Peter Simons, Virginie Bondu, Angela Wandinger-Ness, Tione Buranda [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Small, monomeric guanine triphosphate hydrolases (GTPases) are ubiquitous cellular integrators of signaling. A signal activates th...
[]
83,933
SHARE-seq protocol v2.2
4
null
https://www.protocols.io/view/share-seq-protocol-v2-2-cv75w9q6
Amelia Hall
TITLE: SHARE-seq protocol v2.2 AUTHORS: Amelia Hall [DESCRIPTION] TBD [STEPS] SECTION: 1.1 Ordering Oligo Plates and oligos for plate production 2. One of the more involved aspects of SHARE-seq (and SPLiT-seq) is properly making the oligo hybridization 96 well plates - this section covers that in detail before moving...
["[1.1 Ordering Oligo Plates and oligos for plate production] One of the more involved aspects of SHARE-seq (and SPLiT-seq) is properly making the oligo hybridization 96 well plates - this section covers that in detail before moving into any of the day to day aspects of this protocol (those start at step 24, in section...
33,852
Ferrous Ion-chelating Activity
null
dx.doi.org/10.17504/protocols.io.bda4i2gw
null
Jing Xu
TITLE: Ferrous Ion-chelating Activity AUTHORS: Jing Xu [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>The effects of ferrous ion-chelating activity were conducted according to Guo et al.An aliquot of 1 ml extract was added to a solution of 100 µL of FeCl</span><span style = "vertical-align:su...
[]
30,471
6-Hydroxydopamine (6-OHDA) Treatment and Neurite Tracing in miDA Neurons
1
null
https://www.protocols.io/view/6-hydroxydopamine-6-ohda-treatment-and-neurite-tra-9zfh73n
Yingchao Xue, Xiping Zhan, Shisheng Sun, Senthilkumar S. Karuppagounder, Shuli Xia, Valina L Dawson, Ted M Dawson, John Laterra, Jianmin Zhang, Mingyao Ying
TITLE: 6-Hydroxydopamine (6-OHDA) Treatment and Neurite Tracing in miDA Neurons AUTHORS: Yingchao Xue, Xiping Zhan, Shisheng Sun, Senthilkumar S. Karuppagounder, Shuli Xia, Valina L Dawson, Ted M Dawson, John Laterra, Jianmin Zhang, Mingyao Ying [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>...
["[Cell Seeding]\nPlate miDA neurons (3 × 103 per well) in a 384-well plate coated with poly-D-lysine and laminin.", "[Calcein AM Assay]\nPrepare fresh 6-OHDA in vehicle solution (0.15% ascorbic acid in H2O).\nControl cells should be treated with vehicle solution alone.", "[Calcein AM Assay]\nIncubate cells with fresh ...
26,995
Generation and Purification of pTXB1.Tn5
1
dx.doi.org/10.17504/protocols.io.6kthcwn
https://www.protocols.io/view/generation-and-purification-of-ptxb1-tn5-6kthcwn
Ryan Mulqueen, Andy Fields, Andrew Adey
TITLE: Generation and Purification of pTXB1.Tn5 AUTHORS: Ryan Mulqueen, Andy Fields, Andrew Adey [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Generation of Tn5 transposome, protein purification and loading for the sci- family of protocols.</div><div class = "text-block">Protein purificatio...
["[Prior to day 1]\nGenerate LB agarose plates with following addgene suggested protocol. https://www.addgene.org/mol-bio-reference/#antibiotics\n[Carbenicillin]", "[Day 1: Grow fresh colonies]\nFreshly streak out a LB agarose plates with from the glycerol stock. Allow to grow overnight at in an incubator.\n[Car...
84,606
Roteiro para vídeo-devolutivas de relatórios actigráficos
1
dx.doi.org/10.17504/protocols.io.261gedozdv47/v1
https://www.protocols.click/view/roteiro-para-v-deo-devolutivas-de-relat-rios-actig-cwu6xeze
Daniel Vartanian, Alicia Rafaelly Vilefort Sales, Maria Augusta Medeiros Andrade
TITLE: Roteiro para vídeo-devolutivas de relatórios actigráficos AUTHORS: Daniel Vartanian, Alicia Rafaelly Vilefort Sales, Maria Augusta Medeiros Andrade [DESCRIPTION] Este protocolo foi desenhado para o processo de coleta de dados do projeto de pesquisa Sono e Gravidez. Consulte a aba Guidelines para visualizar o fl...
["[Preparação] Abra o relatório actigráfico em uma janela única em seu navegador.", "[Preparação] Oculte o menu do Google Docs ao clicar na seta localizada no canto superior direito da página.", "[Preparação] Caso alguém, além de você, esteja visualizando o documento, peça para que eles saiam.", "[Preparação] Abra o Lo...
49,737
Discovery proteomic (DDA) LC-MS/MS data acquisition and analysis
1
dx.doi.org/10.17504/protocols.io.buthnwj6
https://www.protocols.io/view/discovery-proteomic-dda-lc-ms-ms-data-acquisition-buthnwj6
Yan Chen, Jennifer Gin, Christopher Petzold
TITLE: Discovery proteomic (DDA) LC-MS/MS data acquisition and analysis AUTHORS: Yan Chen, Jennifer Gin, Christopher Petzold [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol details steps in discovery proteomic data-dependent acquisition with a standard-flow UHPLC-Obitrap system a...
["[Proteomics: HPLC and Mass Spectromtery]\nThaw peptide samples , and transfer of each sample to LC autosampler vials (Agilent, Cat.#5182-0567,#5182-0564) or 96-well plate (Bio-Rad, Cat.#HSP9655).\non ice\n30 µl", "[Proteomics: HPLC and Mass Spectromtery]\nLiquid chromatography-tandem mass spectrometry (LC-MS/MS) an...
74,988
P0 PLANIFICACIÓN ANUAL
1
dx.doi.org/10.17504/protocols.io.5qpvorpb7v4o/v1
https://www.protocols.io/view/p0-planificaci-n-anual-cmgku3uw
cgarcia
TITLE: P0 PLANIFICACIÓN ANUAL AUTHORS: cgarcia [DESCRIPTION] Cada curso académico, la EIDUCAM funcionará con un calendario establecido, que se desarrollará en el mes de Mayo del curso anterior. Fechas a fijar para cada curso académico [STEPS] SECTION: P0 PLANIFICACIÓN ANUAL 1. Fechas a fijar para cada curso académic...
["[P0 PLANIFICACIÓN ANUAL] Fechas a fijar para cada curso académico:\n\nDifusión de la información relativa a los programas de doctorado de la EIDUCAM: Mes de Junio.", "[P0 PLANIFICACIÓN ANUAL] Planificación Programas de Doctorado (Septiembre)\n\na. Oferta de plazas por línea de investigación \nb. Oferta de plazas espe...
86,891
Can light be used to treat obesity and diabetes?
4
dx.doi.org/10.17504/protocols.io.j8nlkooj1v5r/v3
https://www.protocols.io/view/can-light-be-used-to-treat-obesity-and-diabetes-cy4jxyun
Rédoane Daoudi
TITLE: Can light be used to treat obesity and diabetes? AUTHORS: Rédoane Daoudi [DESCRIPTION] This document is highly theoretical lab method paper. It may be used to perform thermogenesis from several cells in a living organism instead of only brown adipocytes. We perform thermogenesis from white adipocytes. The purpo...
["[Introduction] This document is highly theoretical lab method paper. It may be used to perform thermogenesis from several cells in a living organism instead of only brown adipocytes. We perform thermogenesis from white adipocytes. The purpose is to uptake the blood glucose and lipids to produce heat and subsequent we...
40,492
Binding properties of Human IgG to immunoglobulin-binding proteins tested by double immunodiffusion (Ouchterlony) technique.
4
dx.doi.org/10.17504/protocols.io.bjskkncw
https://www.protocols.io/view/binding-properties-of-human-igg-to-immunoglobulin-bjskkncw
Angel Justiz-Vaillant
TITLE: Binding properties of Human IgG to immunoglobulin-binding proteins tested by double immunodiffusion (Ouchterlony) technique. AUTHORS: Angel Justiz-Vaillant [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">1. IBP-HRP conjugates can be prepared in the laboratory by the periodate method.</div>...
["The binding of Human IgG to peroxidase-conjugated Immunoglobulin-binding protein (IBP-HRP) including SpA-HRP, SpG-HRP, SpL-HRP, SpLA-HRP, SpAG-HRP, SpLG-HRP, SpLAG-HRP and turtle serum (Sigma-Aldrich) as a negative control are investigated by double immunodiffusion.", "Briefly, 1% agarose gels are prepared and wells...
null
null
null
dx.doi.org/10.17504/protocols.io.h8vb9w6
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?.
[]
48,702
nCoV-2019 sequencing protocol v3 (LoCost) - InvitaeSF v2.0
1
null
https://www.protocols.io/view/ncov-2019-sequencing-protocol-v3-locost-invitaesf-bts6nnhe
Amanda Kahn-Kirby, Lisa Cunden, Bianca Miani [Invitae
TITLE: nCoV-2019 sequencing protocol v3 (LoCost) - InvitaeSF v2.0 AUTHORS: Amanda Kahn-Kirby, Lisa Cunden, Bianca Miani [Invitae [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Amplicon sequencing protocol for SARS-CoV-2 v3 (LoCost), modified for automation and clean yield. Thank you Josh Quick and...
["[cDNA preparation]\nStamp input plate to a 3-col or 6-col pre-aliquoted RT master mix plate using the Bravo protocol. AB1ComponentVolume2LunaScript RT SuperMix (5X) 2 µL3Normalized Template RNA 8 µL4Total10 µL\nAB1ComponentVolume2LunaScript RT SuperMix (5X) 2 µL3Normalized Template RNA 8 µL4Total10 µL\nViral R...
70,678
Isolating bacteria from algal monocultures
4
dx.doi.org/10.17504/protocols.io.261ge3okyl47/v1
https://www.protocols.io/view/isolating-bacteria-from-algal-monocultures-cg9wtz7e
Emily Aguirre
TITLE: Isolating bacteria from algal monocultures AUTHORS: Emily Aguirre [DESCRIPTION] To isolate bacteria from non-axenic, algal (we did this with Symbiodinium but may work with other algae) monocultures. Our aim is to isolate bacterial members of the phycosphere to use in re-inoculation and genome wide association s...
["[Initial isolation from Symbiodinium] Transfer 1 mL of a non-axenic Symbiodinium culture to a 2mL microcentrifuge tube. Centrifuge the algae and bacteria at 10,000 rpm for 2 minutes.", "[Initial isolation from Symbiodinium]", "[Initial isolation from Symbiodinium]", "[Initial isolation from Symbiodinium]", "[Initial ...
45,064
Quick Protocol for Monarch® DNA Gel Extraction Kit (NEB #T1020)
1
dx.doi.org/10.17504/protocols.io.bp9gmr3w
https://www.protocols.io/view/quick-protocol-for-monarch-dna-gel-extraction-kit-bp9gmr3w
New England Biolabs
TITLE: Quick Protocol for Monarch® DNA Gel Extraction Kit (NEB #T1020) AUTHORS: New England Biolabs [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This is the quick version of the Monarch® DNA Gel Extraction Kit Protocol (NEB #T1020). For the full protocol, please click </span><a href="https:...
["Excise the DNA fragment from the agarose gel, taking care to trim excess agarose. Transfer to a 1.5 ml microfuge tube and weigh the gel slice. Minimize exposure to UV light.\nTrim/remove excess agarose and minimize exposure to UV light.", "Add 4 volumes of Gel Dissolving Buffer to the gel slice (e.g., 400 μl buffer p...
108,552
Designing sgRNA Oligos and Inserting Guides into the GEARBOCS Vector
0
dx.doi.org/10.17504/protocols.io.5jyl8271rl2w/v1
https://www.protocols.io/view/designing-sgrna-oligos-and-inserting-guides-into-t-dm9g493w
Luke Bradley
TITLE: Designing sgRNA Oligos and Inserting Guides into the GEARBOCS Vector AUTHORS: Luke Bradley [DESCRIPTION] This protocol describes designing sgRNAs for genes of interest using the GEARBOCS vector backbone. This method allows researchers to utilize the GEARBOCS system for astrocyte-specific genetic manipulation i...
["[Designing sgRNA oligos] Use UCSC to get gene's exons and introns. Make sure the area you are targeting is in all isoforms.", "[Designing sgRNA oligos] Before designing your own guides, check the literature for guides that you can use.", "[Designing sgRNA oligos] Second, try CRISPick with the exon you want to target....
89,164
Wnt-3a and R-spo1 conditioned media reporter assay
4
dx.doi.org/10.17504/protocols.io.5qpvorz99v4o/v2
https://www.protocols.io/view/wnt-3a-and-r-spo1-conditioned-media-reporter-assay-c3bkyikw
Natalia Martagón, Gurdrun Kliem
TITLE: Wnt-3a and R-spo1 conditioned media reporter assay AUTHORS: Natalia Martagón, Gurdrun Kliem [DESCRIPTION] Protocol designed to measure the activity of Wnt-3a or R-spondin-1 (Rspo1) conditioned media. A reporter HEK cell line expressing luciferase under Wnt-3a stimulation is cultured with conditioned media follo...
["[Day 2: cell stimulation] Aspirate medium and discard.\n\nAdd a total of 500 µL of HEK medium with the following amount of conditioned medium (CM) to test:\nFor R-spondin CM: 12.5 % volume Wnt-3a CM + 2.5 % volume Rspo1 CM (3)\nFor Wnt3a CM .50 % volume", "[Day 1: Seeding of Hek 293 STF cells] Start with one almost...
65,633
Development of a simple and versatile in vitro method for production, stimulation, and analysis of bioengineered muscle
4
dx.doi.org/10.17504/protocols.io.ccb9ssr6
https://www.protocols.io/view/development-of-a-simple-and-versatile-in-vitro-met-ccb9ssr6
Karen Wells-Cembrano, Júlia Sala, Jose Antonio del Rio
TITLE: Development of a simple and versatile in vitro method for production, stimulation, and analysis of bioengineered muscle AUTHORS: Karen Wells-Cembrano, Júlia Sala, Jose Antonio del Rio [DESCRIPTION] In recent years, 3D in vitro modeling of human skeletal muscle has emerged as a subject of increasing interest, du...
["[Making muscle culture devices] Mark a line of 1.2 cm in the center of each 35 mm ∅ Petri dish. In order to obtain consistently centered markings, it is recommended to make a small stencil out of paper or plastic (Figure 2).", "[Preparation of cells and materials] Defrost myoblast cell line in a T75 flask and culture...
49,548
SARS-CoV2 GISAID submission protocol
1
dx.doi.org/10.17504/protocols.io.bumknu4w
https://www.protocols.io/view/sars-cov2-gisaid-submission-protocol-bumknu4w
Nabil-Fareed Alikhan, Emma Griffiths, Ruth Timme, Duncan MacCannell
TITLE: SARS-CoV2 GISAID submission protocol AUTHORS: Nabil-Fareed Alikhan, Emma Griffiths, Ruth Timme, Duncan MacCannell [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol provides the steps needed to establish a new GISAID submission environment for your laboratory. Once established, th...
["[GISAID Submissions]\nGISAID submissions require contextual information (metadata) and the COVID-19 consensus sequence or genome sequence. It is much simpler than other databases, as it has no hierachical structure. In GISAID, one set of contextual information links to one genome assembly.We advise that contextual in...
63,523
https://www.facebook.com/Condor-CBD-Gummies-104105128992798
1
dx.doi.org/10.17504/protocols.io.eq2lyn2jrvx9/v1
https://www.protocols.io/view/https-www-facebook-com-condor-cbd-gummies-10410512-caabsaan
Condor-CBD-Gummies
TITLE: https://www.facebook.com/Condor-CBD-Gummies-104105128992798 AUTHORS: Condor-CBD-Gummies [DESCRIPTION] to assist you with recuperating quickly and regularly with the top promoting hemp oil confine! This uncommon color utilizes 100% regular dynamic fixings just as NO THC to ensure that you get your preeminent rec...
["Condor CBD Gummies\n\n \n\n\n➢Product Name —Condor CBD Gummies Reviews\n➢Main Benefits—Improve Metabolism & Help in Pain Relief\n➢Composition —NaturalOrganic Compound\n➢Side-Effects—NA\n➢Rating :—⭐⭐⭐⭐⭐\n➢Availability —Online\n➢Price (for Sale) Buy Now Here —Click Here\nCondor CBD Gummies - You plan to recuperate just...
44,740
Geometric Method for Estimating Coral Surface Area Using Image Analysis
1
dx.doi.org/10.17504/protocols.io.bpxcmpiw
https://www.protocols.io/view/geometric-method-for-estimating-coral-surface-area-bpxcmpiw
Rowan Mclachlan, Andrea Grottoli
TITLE: Geometric Method for Estimating Coral Surface Area Using Image Analysis AUTHORS: Rowan Mclachlan, Andrea Grottoli [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol outlines a non-destructive geometric method for estimating the surface area of Scleractinian coral samples with relat...
["[Photographing corals]\nPrepare the photograph area. Setup a tripod so that the digital camera is pointing downward, parallel to the workbench as shown in Fig. 1. Place a metric ruler on two blocks of equal height (at least two inches) and make sure a section of the ruler is visible in the digital camera field of vie...
66,323
Cannaleafz CBD Gummies Canada Reviews: Shark Tank Ingredients &amp; Price on Website
3
dx.doi.org/10.17504/protocols.io.eq2lynrpevx9/v1
https://www.protocols.io/view/cannaleafz-cbd-gummies-canada-reviews-shark-tank-i-ccztsx6n
Cannaleafz CBD Gummies
TITLE: Cannaleafz CBD Gummies Canada Reviews: Shark Tank Ingredients &amp; Price on Website AUTHORS: Cannaleafz CBD Gummies [DESCRIPTION] Cannaleafz CBD Gummies Reviews:CBD is generally utilized in different nations of the world for restorative purposes. Be that as it may, many individuals get high in the wake of con...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.estbeen
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>PBCV-1 is a large plaque virus (approximately 2-3 mm in diameter).  The plaque size of the viruses reflects the length of the virus replication cycle.  PBCV-1 has a life cycle of about 6-8 hours.  When titering you should use plates that were poured no more than 1 day before ...
[]
82,033
Transferring C. elegans to S-basal to grow them in liquid culture
4
dx.doi.org/10.17504/protocols.io.81wgbyj41vpk/v1
https://www.protocols.io/view/transferring-c-elegans-to-s-basal-to-grow-them-in-cucrwsv6
Muhammad Zaka Asif, Man Shah, Yosef Smadi
TITLE: Transferring C. elegans to S-basal to grow them in liquid culture AUTHORS: Muhammad Zaka Asif, Man Shah, Yosef Smadi [DESCRIPTION] This protocol describes day 2 of our workflow to grow worms in liquid culture to induce the production of natural products (in this case, 1-HP derivatives). In this protocol, we get...
["Remove flasks with L1-arrested worms from the shaker", "Swirl vigorously for at least 30 seconds, then collect a 1 uL aliquot and place onto a glass slide for inspection. Use a 0.5 uL aliquot if there are more than 100 worms in the 1 uL aliquot.", "Count using a tally counter. Repeat for at least 3 trials (may need t...
null
null
null
dx.doi.org/10.17504/protocols.io.mk5c4y6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>In this protocol,<span lang="en"><span class=""> the extraction of template DNA out of <em>Synechocystis</em> sp. PCC 6803 samples, the PCR of the template and the application on a agarosegel are described. The protocol was handed over by Maximilian Dietsch MSc.<br /></span><...
[]
66,983
Ice House Portable AC
3
dx.doi.org/10.17504/protocols.io.n92ldzdmov5b/v1
https://www.protocols.io/view/ice-house-portable-ac-cdnfs5bn
hggsasd
TITLE: Ice House Portable AC AUTHORS: hggsasd [DESCRIPTION] Ice House Portable AC [STEPS]
[]
103,473
Nuclei Isolation from Human Subchondral Bone for 10x Multiome
0
dx.doi.org/10.17504/protocols.io.j8nlk8mrdl5r/v1
https://www.protocols.io/view/nuclei-isolation-from-human-subchondral-bone-for-1-dhar32d6
Takuya Sakamoto, Merissa Olmer, Martin Lotz
TITLE: Nuclei Isolation from Human Subchondral Bone for 10x Multiome AUTHORS: Takuya Sakamoto, Merissa Olmer, Martin Lotz [DESCRIPTION] This protocol describes isolation of nuclei from fresh-frozen human knee subchondral bone for use in Omics analyses, including RNA-sequencing of ATAC-sequencing. Tissue dissociation w...
[]
64,826
Keto Complete ( Fraud Attention 2022 ) - Do Really Work Keto Complete Australia?
3
dx.doi.org/10.17504/protocols.io.rm7vzy918lx1/v1
https://www.protocols.io/view/keto-complete-fraud-attention-2022-do-really-work-cbi2skge
jackweyand
TITLE: Keto Complete ( Fraud Attention 2022 ) - Do Really Work Keto Complete Australia? AUTHORS: jackweyand [DESCRIPTION] Keto Complete Australia [STEPS]
[]
106,299
SEQUENCING Protocol Template
1
null
https://www.protocols.io/view/sequencing-protocol-template-dj234qgn
Kathleen Pitz, Raïssa Meyer
TITLE: SEQUENCING Protocol Template AUTHORS: Kathleen Pitz, Raïssa Meyer [DESCRIPTION] A protocol template created through the BeBOP project for sequencing. Main protocol steps begin below the "Standard Operating Procedure" section. A detailed description of the BeBOP project and these templates is available here: ht...
["[MIOP: Minimum Information about an Omics Protocol] MIOP Term\nValue\n \nmethodology category\n\n \nproject\n\n \npurpose\n\n \nanalyses\n\n \ngeographic location\n\n \nbroad-scale environmental context\n\n \nlocal environmental context\n\n \nenvironmental medium\n\n \ntarget\n\n \ncreator\n\n \nmaterials required\...
80,834
Laboratory and radiological data collection
1
dx.doi.org/10.17504/protocols.io.6qpvr459ogmk/v1
https://www.protocols.io/view/laboratory-and-radiological-data-collection-cs7awhie
Kajiru Gad Kilonzo, Stefanie J. Krauth, Jo Halliday, Clive Kelly, Stefan Siebert, Gloria Temu, Christopher Bunn, Nateiya M Yongolo, Sally Wyke, Emma McIntosh, Blandina Mmbaga
TITLE: Laboratory and radiological data collection AUTHORS: Kajiru Gad Kilonzo, Stefanie J. Krauth, Jo Halliday, Clive Kelly, Stefan Siebert, Gloria Temu, Christopher Bunn, Nateiya M Yongolo, Sally Wyke, Emma McIntosh, Blandina Mmbaga [DESCRIPTION] This protocol details laboratory and radiological data collection. [S...
["[Laboratory and radiological data collection] Following completion of the screening and examinations within both the community and hospital data capture, adult patients with evidence of synovitis or joint swelling will have 20 mL blood taken for later analysis of acute phase reactants (ESR, CRP) and autoantibodies (R...
null
null
null
dx.doi.org/10.17504/protocols.io.dxb7im
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?.
[]
60,213
eDNA extraction with kaneka
1
dx.doi.org/10.17504/protocols.io.dm6gpb9xplzp/v1
https://www.protocols.io/view/edna-extraction-with-kaneka-b62vrge6
Fei Xia
TITLE: eDNA extraction with kaneka AUTHORS: Fei Xia [DESCRIPTION] We used the Kaneka Simple DNA Extraction Kit version 2 (Kaneka) for on-site DNA extraction from Sterivex. We injected 500 µl of Solution A of the kit into the Sterivex and shook it by hand for 1 min. We collected the Solution A buffer in the Sterivex b...
["Inject 250 µL of A buffer into the Sterivex, 1 min shake and rotate.", "Put Sterivex into 50ml tube and secured for centrifugation 5000 rpm, 5 min", "Add 40 µL of B buffer to the 1.5ml tube.", "Store the samples at -20 °C .", "Remove the liquid from the inlet and put into the 1.5ml tube.", "Mix by vortexing."]
59,225
Small Object and Artefact Photography - &#39;SOAP&#39; Protocol
5
dx.doi.org/10.17504/protocols.io.b53zq8p6
https://www.protocols.io/view/small-object-and-artefact-photography-39-soap-39-p-b53zq8p6
Jacopo Niccolo Cerasoni, Felipe Do Nascimento Rodrigues
TITLE: Small Object and Artefact Photography - &#39;SOAP&#39; Protocol AUTHORS: Jacopo Niccolo Cerasoni, Felipe Do Nascimento Rodrigues [DESCRIPTION] Photography is among one of the most widely used methods in scientific publications to efficiently and objectively communicate morphological, technological and aesthet...
["[Collect Equipment] Collect and prepare equipment (see materials section for full list of equipment).", "[Collect Equipment] Arguably the most important part of a camera is the lens. Before continuing with this method, we suggest the reader understand what type of lenses exist and which one works best for its use. In...
63,158
Keto Blast Gummies Reviews &amp; Shark Tank 2022
1
dx.doi.org/10.17504/protocols.io.6qpvr67q2vmk/v1
https://www.protocols.io/view/keto-blast-gummies-reviews-amp-shark-tank-2022-b9wwr7fe
ketoblastiesreviews
TITLE: Keto Blast Gummies Reviews &amp; Shark Tank 2022 AUTHORS: ketoblastiesreviews [DESCRIPTION] Keto Blast Gummies Reviews & Shark Tank 2022 [STEPS] 1. Keto Blast Gummies Reviews & Shark Tank 2022 Kwazi Keto Gummies & Keto Blast Gummies As we know that weight loss is a lengthy process. It isn't possible to lose...
["Keto Blast Gummies Reviews & Shark Tank 2022\nKwazi Keto Gummies & Keto Blast Gummies As we know that weight loss is a lengthy process. It isn't possible to lose a lot of weight in one day. Numerous people tried their stylish to get into the right shape but they didn’t succeed. On the other hand, exercise and diet pl...
28,348
Naegleria gruberi plasmids
null
dx.doi.org/10.17504/protocols.io.7w4hpgw
https://www.protocols.io/view/naegleria-gruberi-plasmids-7w4hpgw
Anastasios Tsaousis, Eleana Kazana, Tobias von der Haar
TITLE: Naegleria gruberi plasmids AUTHORS: Anastasios Tsaousis, Eleana Kazana, Tobias von der Haar [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Maps and nucleotide sequences of the Naegleria gruberi transfection plasmids (pNaeg-HYG and pNaeg-NEO) reported in Faktorová D. et al. </div></div> [STE...
[]
71,066
Sample preconditioning before scanning electron microscopy (SEM)
4
dx.doi.org/10.17504/protocols.io.36wgqj465vk5/v1
https://www.protocols.io/view/sample-preconditioning-before-scanning-electron-mi-chm2t48e
An.Huang
TITLE: Sample preconditioning before scanning electron microscopy (SEM) AUTHORS: An.Huang [DESCRIPTION] Bacterial samples need to be preconditioned before visualized by scanning electron microscope (SEM). This protocol describes preconditioning methods of bacteria before SEM. [STEPS] SECTION: Bacterial samples precon...
["[Bacterial samples preconditioning] Obtain a small quantity of bacteria to be tested from a culture.", "[Bacterial samples preconditioning] Centrifuge the samples at5000 rpm, 10 min, 4 °C . Resuspend the pellet using 1X phosphate buffed saline (PBS). Repeat this step for three times. Keep the supernatant after the th...
29,947
High molecular weight DNA extraction from fungal tissue for the long read PacBio sequencing
null
dx.doi.org/10.17504/protocols.io.9g3h3yn
null
Reynaldi Darma, Megan McDonald
TITLE: High molecular weight DNA extraction from fungal tissue for the long read PacBio sequencing AUTHORS: Reynaldi Darma, Megan McDonald [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Obtaining high molecular weight DNA from certain materials for PacBio sequencing can be quite challenging because...
["[Grinding the spores]\nGrind fungal tissues 3-4 times for 10-15 sec bursts adding liquid nitrogen each time. Do not exert excessive pressure while grinding the spores as this may shear the DNA.", "[DNA extraction]\nAdd 200 µl proteinase Proteinase K (20 mg/mL) and incubate at RT for 1.5 hours on the orbital shaker wi...
98,034
FUNDIS version ONT dA-tailing for Fungal Barcoding
4
null
https://www.protocols.io/view/fundis-version-ont-da-tailing-for-fungal-barcoding-dbys2pwe
Stephen Douglas Russell, Harte Singer
TITLE: FUNDIS version ONT dA-tailing for Fungal Barcoding AUTHORS: Stephen Douglas Russell, Harte Singer [DESCRIPTION] This protocol is for dA-tailing, which is an enzymatic method for adding a non-templated nucleotide to the 3' end of a blunt, double-stranded DNA molecule. In other words, this puts A-chains on the en...
["[End repair/A-tailing] Put a 1.5mL aliquot of fresh molecular water on a heat block at 55 °C. This will be used after the cleanup step towards the end of this protocol.\n\nTurn on your PCR thermal cycler so that the heated lid begins to come up to temp.\n\nTake out the materials for library prep from the freezer. Fli...
88,107
Ethnic-Racial Socialization in Early Childhood: Scoping Review Protocol
1
dx.doi.org/10.17504/protocols.io.q26g7pmq8gwz/v1
https://www.protocols.io/view/ethnic-racial-socialization-in-early-childhood-sco-c2ajyacn
Erica E Coates, Charles S. Dorris MLIS, Alison B McLeod, Kaela Farrise
TITLE: Ethnic-Racial Socialization in Early Childhood: Scoping Review Protocol AUTHORS: Erica E Coates, Charles S. Dorris MLIS, Alison B McLeod, Kaela Farrise [DESCRIPTION] Objective: This scoping review aims to assess the state of the literature on ethnic-racial socialization (ERS) during the early childhood period. ...
["[Introduction] Caregivers use ethnic-racial socialization (ERS) to transmit information about race and ethnicity to their children (Hughes et al., 2006). Research on ERS has increased over the past 20 years but has mostly focused on ERS with older children and adolescents (see Umaña‐Taylor & Hill, 2020 for a review)....
71,152
Manual Blood Pressure (Auscultation Technique)
1
dx.doi.org/10.17504/protocols.io.5jyl8j568g2w/v1
https://www.protocols.click/view/manual-blood-pressure-auscultation-technique-chqqt5vw
g.spencer
TITLE: Manual Blood Pressure (Auscultation Technique) AUTHORS: g.spencer [DESCRIPTION] The following SOPs outline how to take a blood pressure measurement both manually and using an automated device. Blood-pressure measurement is warranted in any situation that requires assessment of cardiovascular health, including ...
["[Equipment Required] Stethoscope\nSphygmomanometer (cuff, bulb and a manometer)", "[Preparing for the Measurement] Follow the Steps below to help prepare your PPC for measurement:\n\nThe patient’s back and legs should be supported with legs uncrossed and feet resting on a firm surface\n\nThe patient’s arms should be ...
null
null
null
dx.doi.org/10.17504/protocols.io.iqqcdvw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Forms used for data collection in the study entitled: “Risk factors for adverse drug reactions in pediatric inpatients: a cohort study”</p> <p> </p> <p>Forms 1 and 2 must be used in the first pharmaceutical consultation after hospital admission of the patient:</p> <p>i) form ...
[]
56,735
spyDNA650 staining with CO2 independent media for live endothelium microscopy
4
null
https://www.protocols.io/view/spydna650-staining-with-co2-independent-media-for-b3m7qk9n
Emir Bora Akmeriç
TITLE: spyDNA650 staining with CO2 independent media for live endothelium microscopy AUTHORS: Emir Bora Akmeriç [DESCRIPTION] "spyDNA650 staining with CO2 independent media for live endothelium microscopy" [STEPS] SECTION: Staining 1. Dilute 1:1000 SPY650 DNA in CO2 independent media(500 uL total volume for 2 slide...
["[Staining] Dilute 1:1000 SPY650 DNA in CO2 independent media(500 uL total volume for 2 slides, 220 uL per slide and incubate in incubator for 20 mins", "[Staining] Gently remove overnight media from the u-slides and add warmed dye-media mix. Check whether monolayer is preserved. incubate for 2-3 hours", "[Flow Unit E...
73,157
Adquisición Desideratas Biblioteca UCAM
1
dx.doi.org/10.17504/protocols.io.81wgbyzdyvpk/v1
https://www.protocols.io/view/adquisici-n-desideratas-biblioteca-ucam-cjpdumi6
Antonio Rex Alegria
TITLE: Adquisición Desideratas Biblioteca UCAM AUTHORS: Antonio Rex Alegria [DESCRIPTION] Procedimiento para la Adquisición de material bibliográfico [STEPS] 1. Ve a la página web de la Biblioteca de la UCAM https://www.ucam.edu/biblioteca 2. Pulsa en el icono de Adquisiciones 3. Se abre una nueva pantalla...
["Ve a la página web de la Biblioteca de la UCAM https://www.ucam.edu/biblioteca", "Pulsa en el icono de Adquisiciones", "Se abre una nueva pantalla.", "Seleccionamos el icono de Solicitud de Adquisiciones/Soporte.", "Nos identificamos con nuestro usuario y contraseña.", "Introducimos los datos que nos solicita el sop...
61,814
LDM protocol for estimating plasmid conjugation rates
4
null
https://www.protocols.io/view/ldm-protocol-for-estimating-plasmid-conjugation-ra-b8kwruxe
Olivia Kosterlitz, Claire Wate
TITLE: LDM protocol for estimating plasmid conjugation rates AUTHORS: Olivia Kosterlitz, Claire Wate [DESCRIPTION] This is a detailed protocol to implement the LDM approach for estimating plasmid conjugation rates. To get a conceptual overview of the approach, please read the accompanying manuscript: The protoco...
["[Phase 1: Minimum inhibitory concentration assay with the transconjugant-selecting medium. medium.] Prepare bacterial cultures.\n\n Start cultures from freezer stocks of the donors, recipients, and transconjugants. Supplement the donor and transconjugant growth culture medium with the appropriate selection to maintai...
74,910
iPad_Lidar_surface_roughness_processing
5
dx.doi.org/10.17504/protocols.io.6qpvr4pe3gmk/v1
https://www.protocols.io/view/ipad-lidar-surface-roughness-processing-cmd6u29e
Alex White
TITLE: iPad_Lidar_surface_roughness_processing AUTHORS: Alex White [DESCRIPTION] This protocol was used to process LIDAR data acquired over 3-m transects of forest floor, using an Apple iPad Pro 2022 app entitled 3D Scanner App in “LIDAR Advanced” mode set to the smallest possible maximum depth (30 cm) and resolution ...
["Open CloudCompare.", "Open textured output file:", "Select the file in the DB Tree on the left. \n \n A bounding box will appear around the mesh.", "Click the Cross Section icon in the toolbar.", "Click the folder icon.", "Navigate to a transect folder (e.g. MA404-1-EW_20_02_48) in Box > SMAPVEX22_data > MA_lid...
null
null
null
dx.doi.org/10.17504/protocols.io.kq9cvz6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>The appropriate use of clinically accurate diagnostic tests is essential for the detection of pertussis, a poorly controlled vaccine-preventable disease. The purpose of this study was to estimate the sensitivity and specificity of different diagnostic criteria including cultu...
[]
44,323
Quantitate Library
4
dx.doi.org/10.17504/protocols.io.bpibmkan
https://www.protocols.io/view/quantitate-library-bpibmkan
Eric L. Van Nostrand, Thai B. Nguyen, Chelsea Gelboin-Burkhart, Ruth Wang, Steven M. Blue, Gabriel A. Pratt, Ashley L. Louie, Gene W. Yeo
TITLE: Quantitate Library AUTHORS: Eric L. Van Nostrand, Thai B. Nguyen, Chelsea Gelboin-Burkhart, Ruth Wang, Steven M. Blue, Gabriel A. Pratt, Ashley L. Louie, Gene W. Yeo [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Profiling of RNA binding protein targets in vivo provides critical insights int...
["Quantitate using D1000 DNA Tapes for the Agilent TapeStation."]
76,026
RCA-NGS for RNA viruses
4
dx.doi.org/10.17504/protocols.io.rm7vzb7wrvx1/v1
https://www.protocols.io/view/rca-ngs-for-rna-viruses-cng2vbye
Masayasu Misu, Tomoki Yoshikawa, Satoko Sugimoto, Yuki Takamatsu, Takeshi Kurosu, Yukiteru Ouji, Masahide Yoshikawa, Masayuki Shimojima, Hideki Ebihara, Masayuki Saijo
TITLE: RCA-NGS for RNA viruses AUTHORS: Masayasu Misu, Tomoki Yoshikawa, Satoko Sugimoto, Yuki Takamatsu, Takeshi Kurosu, Yukiteru Ouji, Masahide Yoshikawa, Masayuki Shimojima, Hideki Ebihara, Masayuki Saijo [DESCRIPTION] This RCA-NGS were optimized for an NGS machine, MinION. These methods do not require nucleic acid...
["[Preparation for virus supernatant] Centrifuge the working stock virus to remove debris.\n 6000 x g, 10 min", "[Preparation for virus supernatant] Unwanted DNA and RNA mainly originating from the virus-infected cells are digested using .", "[The viral genomic RNA extraction] The viral genomic RNA extraction is perfo...
22,292
AFM tip functionalization with glutaraldehyde
null
dx.doi.org/10.17504/protocols.io.zzuf76w
null
George Lykotrafitis
TITLE: AFM tip functionalization with glutaraldehyde AUTHORS: George Lykotrafitis [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Attach ligand protein to the AFM cantilever tip through functionalization. Further use it in the single molecule force spectroscopy to detect corresponding receptors on t...
["Silanize the tip with 2% v/v 3-aminopropyltiethoxysilane (APTES) in acetone for 10 min at 25°C.", "Rinse the tip with deionized (DI) water.", "Immerse the tip in 0.5% v/v glutaraldehyde in DI water for 30 min at 25°C.", "Rinse the tip with DI water.", "Incubate the tip in protein solution (100-300 µg/mL, eg. 100µg/mL...
null
null
null
dx.doi.org/10.17504/protocols.io.cq9vz5
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?.
[]
45,785
A New Absolute Quantification Method for Protein
4
null
https://www.protocols.io/view/a-new-absolute-quantification-method-for-protein-bqxzmxp6
181830691
TITLE: A New Absolute Quantification Method for Protein AUTHORS: 181830691 [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Absolute quantification method is essential for biological research. For absolute quantification, it is usually necessary to draw standard curves according to molecules w...
[]
84,558
Protocol for Identifying Highly Pathogenic Salmonella Using the HPS Multiplex PCR Assay
4
dx.doi.org/10.17504/protocols.io.6qpvr3on3vmk/v1
https://www.protocols.io/view/protocol-for-identifying-highly-pathogenic-salmone-cwtnxeme
Dayna Harhay, Kerry D. Brader, Jim Bono, Gregory P Harhay, Mick Bosilevac, Tatum S Katz
TITLE: Protocol for Identifying Highly Pathogenic Salmonella Using the HPS Multiplex PCR Assay AUTHORS: Dayna Harhay, Kerry D. Brader, Jim Bono, Gregory P Harhay, Mick Bosilevac, Tatum S Katz [DESCRIPTION] This protocol describes how to perform a multiplex PCR assay for the identification of Highly Pathogenic Salmonel...
["[Bacterial Lysis (BAX lysis) Procedure to prepare DNA template] Reaction: \nAdd 100 µL Protease/Lysis Buffer per well in the PCR plate\nAdd 2.5 µL Sample (overnight culture) per well", "[Bacterial Lysis (BAX lysis) Procedure to prepare DNA template] In a PCR plate, dispense 100 µL of the BAX lysis buffer per well, ad...
24,047
Biochemical Measures of Neuropathy - GSSG (Non-Enzymatic)
null
dx.doi.org/10.17504/protocols.io.3qpgmvn
null
Eva Feldman
TITLE: Biochemical Measures of Neuropathy - GSSG (Non-Enzymatic) AUTHORS: Eva Feldman [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span></div><div class = "text-block">Oxidative stress is highly correlated with the metabolic changes caused by hyperglyce...
["[Sample Preparation:]\nPrior to dissection, perfuse tissue with PBS pH7.4 with 0.16 mg/ml heparin.1. If using extract from GSH add 100 μl 0.04 M N-ethylmaleimide to 200 μl homogenate then skip to Performing the Assay or2. Label 3 sets 1.5 ml and 1 set of 0.6 ml micro centrifuge tubes.3. Prepare 3 M HClO4 and freeze....
91,086
Synthesis of Carbon Dots using Citric acid (Solvent Free reaction)
6
dx.doi.org/10.17504/protocols.io.e6nvwd25dlmk/v1
https://www.protocols.io/view/synthesis-of-carbon-dots-using-citric-acid-solvent-c47nyzme
qurashiubaid
TITLE: Synthesis of Carbon Dots using Citric acid (Solvent Free reaction) AUTHORS: qurashiubaid [DESCRIPTION] Carbon Dots are the fluorescent nanoparticles that have wide range of applications. Facile synthesis of Carbon Dots paves the way for making this material mainstream for use. Synthesis of Carbon Dots by using ...
["[Synthesis of Carbon Dots using Citric acid (Solvent Free reaction)] 2 g 120 °C", "[Synthesis of Carbon Dots using Citric acid (Solvent Free reaction)] Once you get the yellow molten compound , drop it in the 0.1 Mass Percent 100ml solution, dropwise and stir it on magnetic stirrer.", "[Synthesis of Carbon Dots usi...
98,185
Single-cell RNA-seq
0
null
https://www.protocols.io/view/single-cell-rna-seq-db5h2q36
Siwei Li
TITLE: Single-cell RNA-seq AUTHORS: Siwei Li [DESCRIPTION] Single-cell RNA-seq [STEPS] SECTION: Single-cell RNA-seq 1. Tissues were transferred to sterile culture dishes with 10 ml of 1× Dulbecco's phosphate-buffered saline (DPBS; Thermo Fisher, Cat. no. 14190144) on ice to remove residual tissue storage solution and...
["[Single-cell RNA-seq] Tissues were transferred to sterile culture dishes with 10 ml of 1× Dulbecco's phosphate-buffered saline (DPBS; Thermo Fisher, Cat. no. 14190144) on ice to remove residual tissue storage solution and then minced on ice. We used the dissociation enzyme 0.25% trypsin (Thermo Fisher, Cat. no. 25200...
47,843
Protocol for SCV-2000bp: a primer panel for SARS-CoV-2 full-genome sequencing.v3.Neb
4
dx.doi.org/10.17504/protocols.io.bsybnfsn
https://www.protocols.io/view/protocol-for-scv-2000bp-a-primer-panel-for-sars-co-bsybnfsn
Valeriia Kaptelova, Speranskaya AS, et.al
TITLE: Protocol for SCV-2000bp: a primer panel for SARS-CoV-2 full-genome sequencing.v3.Neb AUTHORS: Valeriia Kaptelova, Speranskaya AS, et.al [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Here we present a new primer panel that allows amplifying the complete genome of SARS-CoV-2 (the causative vi...
["[1. cDNA]\nRNA samples should be stored at -80 °C and thawed on ice.NGS library preparation and sequencing:cDNAReverse transcription reaction was performed using 10 μL of the RNA samples, random hexanucleotide primers, and Reverta-L kit (AmpliSens, Russia) according to the manufacturer’s instructions. Prepare a ready...
null
null
null
dx.doi.org/10.17504/protocols.io.meyc3fw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This study aimed to investigate the effects of lactic acid bacterial fermentation extract (LABEF) on cystitis induced by <em>Escherichia coli</em> lipopolysaccharide (LPS) in the mouse bladder by pathological analyses and measurement of the levels of tumor necrosis factor-alp...
[]
57,744
Freezing of hPSCs grown on MEFs
4
dx.doi.org/10.17504/protocols.io.b4mqqu5w
https://www.protocols.io/view/freezing-of-hpscs-grown-on-mefs-b4mqqu5w
Hanqin Li, Oriol Busquets, Steven Poser, Dirk Hockemeyer, Frank Soldner
TITLE: Freezing of hPSCs grown on MEFs AUTHORS: Hanqin Li, Oriol Busquets, Steven Poser, Dirk Hockemeyer, Frank Soldner [DESCRIPTION] This protocol describes the standard procedure of freezing human pluripotent stem cells (hPSCs), which were grown on inactivated mouse embryonic fibroblasts (MEFs). General notes 1. T...
["[A. Freezing of hPSCs as single cell solution using trypsin] When hPSCs reach 50% confluency (usually on day 6 from last passage), change medium to 3 ml hPSCs medium + Rock inhibitor, to prepare for freezing the next day.", "[A. Freezing of hPSCs as single cell solution using trypsin] Before starting:\na. Prepare Fre...
96,667
Protocol: Neurophysiological Effects of Craniosacral Treatment on Heart Rate Variability
0
dx.doi.org/10.17504/protocols.io.4r3l22xjjl1y/v1
https://www.protocols.io/view/protocol-neurophysiological-effects-of-craniosacra-dam32c8n
Andrew C Cook, Anna Egli, Nathan E Cohen, Sunni Coyne, Min Chae, Brandon Kapalko, Kyrie Bernardi, Randy Scott DO
TITLE: Protocol: Neurophysiological Effects of Craniosacral Treatment on Heart Rate Variability AUTHORS: Andrew C Cook, Anna Egli, Nathan E Cohen, Sunni Coyne, Min Chae, Brandon Kapalko, Kyrie Bernardi, Randy Scott DO [DESCRIPTION] Craniosacral treatment is an osteopathic technique grounded in the assumption that ther...
["[Title] Neurophysiological Effects of Craniosacral Treatment on Heart Rate Variability", "[Registration] Protocols.io", "[Authors] Andrew C. Cook: Lake Erie College of Osteopathic Medicine - Bradenton; Bradenton, FL\nACook28563@med.lecom.edu \nORCID: https://orcid.org/0000-0001-6332-1993\n\nAnna Egli: Lake Erie Colle...
89,791
The Bcc qPCR NAD assay for the specific rapid quantitative detection of all Bcc species
1
dx.doi.org/10.17504/protocols.io.4r3l223wql1y/v1
https://www.protocols.io/view/the-bcc-qpcr-nad-assay-for-the-specific-rapid-quan-c3w7yphn
Huong Thu Duong, Shannon Fullbrook, Kate Reddington, Elizabeth Minogue, Thomas Barry
TITLE: The Bcc qPCR NAD assay for the specific rapid quantitative detection of all Bcc species AUTHORS: Huong Thu Duong, Shannon Fullbrook, Kate Reddington, Elizabeth Minogue, Thomas Barry [DESCRIPTION] The Bcc qPCR NAD assay presented is an internally controlled duplex assay (incorporating an IAC), targeting a region...
["[Prepare a qPCR master mix (in a DNA-free preparation hood)] Thaw qPCR reagents and samples on the bench, flick to mix and briefly spin down on a tabletop centrifuge.", "[The Bcc qPCR NAD assay] Transfer the plate into the thermal cycler after setting it up appropriately and carry out qPCR reaction at the following c...
57,727
Quantitative detection of vitamin B12 in algae by bioassay and ICP-MS/MS
4
dx.doi.org/10.17504/protocols.io.14egn7726v5d/v1
https://www.protocols.io/view/quantitative-detection-of-vitamin-b12-in-algae-by-b4k7quzn
Sunnyjoy Dupuis, Stefan Schmollinger, Sabeeha S. Merchant
TITLE: Quantitative detection of vitamin B12 in algae by bioassay and ICP-MS/MS AUTHORS: Sunnyjoy Dupuis, Stefan Schmollinger, Sabeeha S. Merchant [DESCRIPTION] This protocol describes two methods for determining the amount of vitamin B12 present in the spent medium and cell lysate of algae cultures. The first meth...
["[E. coli B12 Bioassay]", "[E. coli B12 Bioassay] Prepare cell culture fractions:", "[E. coli B12 Bioassay] Collect 2 mL culture into a 2 ml screw-cap tube.", "[E. coli B12 Bioassay] Centrifuge at >8000 rcf, 2 min. Quickly transfer 950 µL of the spent medium supernatant to each of two 1.5 ml screw-cap tubes without ...
85,883
Mounting and Coverslipping Mouse Brain Sections
1
dx.doi.org/10.17504/protocols.io.n92ldmpy7l5b/v1
https://www.protocols.io/view/mounting-and-coverslipping-mouse-brain-sections-cx43xqyn
Naveen Ouellette, daphne.toglia, Holly Myers
TITLE: Mounting and Coverslipping Mouse Brain Sections AUTHORS: Naveen Ouellette, daphne.toglia, Holly Myers [DESCRIPTION] This protocol details mounting thin sliced mouse brain tissue sections onto charged slides in a uniform orientation that does not create wrinkles or artifacts that might interfere with imaging, an...
["[Mounting] Place a drop of 1xPBS onto a 1\"x3\" slide.", "[Mounting] Using a small paintbrush, lift the first tissue section to be mounted out of the storage well plate and place section into the petri dish so that it lays flat close to the surface of the 1xPBS & Triton X-100 0.2% solution.", "[Mounting] Repeat steps...
108,826
Crystallisation of Enterovirus D68 3C protease
1
dx.doi.org/10.17504/protocols.io.rm7vzjp45lx1/v1
https://www.protocols.io/view/crystallisation-of-enterovirus-d68-3c-protease-dnh25b8e
ryan Lithgo, Peter Marples, Lizbé Koekemoer, Daren Fearon
TITLE: Crystallisation of Enterovirus D68 3C protease AUTHORS: ryan Lithgo, Peter Marples, Lizbé Koekemoer, Daren Fearon [DESCRIPTION] The development of effective broad-spectrum antivirals forms an important part of preparing for future pandemics. A cause for concern is the currently emerging pathogen Enterovirus D68...
["[Crystallisation experiment] Protein and buffer requirements:\n14.4 µL35 mg/mL \n2.88 mL \n7.2 µL seeds, dilution 1:1 000 000", "[Crystallisation experiment] Dispense 30 µL into SwissCI 3 lens plate reservoir wells using a 100 µl multi-channel pipette.\nDispense 50 nL35 mg/mL to each lens using the SPT mosquito.\nDi...
null
null
null
dx.doi.org/10.17504/protocols.io.hhab32e
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <div> <div> <div> <p>This protocol is from 'Flower Ca<sup>2+</sup> channel in CME and ADBE' of Yao CK et al.</p> <div> <div> <p> </p> <p>Please see the manuscript for the full method details.</p> </div> </div> </div> </div> </div> [BEFORE_START] <p>You'll need: </p> <p> </p> <p...
[]
96,702
Bacterial-induced neutrophilic nasal inflammation in mice
0
dx.doi.org/10.17504/protocols.io.rm7vzxwzxgx1/v1
https://www.protocols.io/view/bacterial-induced-neutrophilic-nasal-inflammation-dan62dhe
Alba Sánchez Montalvo, Aaron Ziani Zeryouh, Marylène Lecocq, Charles Pilette, Valérie Hox
TITLE: Bacterial-induced neutrophilic nasal inflammation in mice AUTHORS: Alba Sánchez Montalvo, Aaron Ziani Zeryouh, Marylène Lecocq, Charles Pilette, Valérie Hox [DESCRIPTION] The present surgical procedure provides a step-by-step and detailed protocol with all the critical information to develop a bacterial-induced...
["[The day before surgery] Prepare the chosen bacteria", "[Surgical procedure] Shave the snout over the intervention area and fix the mouse on a sterile field (Figure 1, A).", "[The day before surgery] Prepare the nasal tampon", "[Pre-surgery] Weight the mouse and administer general anesthesia accordingly: a mix of Xyl...
78,849
Prueba2
4
dx.doi.org/10.17504/protocols.io.j8nlkwyxdl5r/v1
https://www.protocols.io/view/prueba2-cq89vzz6
Yo
TITLE: Prueba2 AUTHORS: Yo [DESCRIPTION] Resuemem [STEPS] 1. Saludar 2. Preguntar como estas? 3. Decir adios
["Saludar", "Preguntar como estas?", "Decir adios"]
106,016
DNA Extraction from Swabbed Feces for Depositor Species Identification - 2024
0
dx.doi.org/10.17504/protocols.io.5qpvokdn9l4o/v1
https://www.protocols.io/view/dna-extraction-from-swabbed-feces-for-depositor-sp-djr84m9w
Karla Vargas
TITLE: DNA Extraction from Swabbed Feces for Depositor Species Identification - 2024 AUTHORS: Karla Vargas [DESCRIPTION] Protocol developed by the Culver Lab at the University of Arizona to extract DNA from feces for depositor species identification [STEPS] SECTION: DAY 1 2. Saturate cotton-tipped applicators with PB...
["[DAY 1] Saturate cotton-tipped applicators with PBS buffer.\n● Keep cotton-tipped applicators in PBS buffer for at least 3 minutes.", "[DAY 1] Pipette 25 μl of proteinase K and 300 μl of ATL buffer (from DNeasy kit) into a new, labeled 2 ml microcentrifuge tube.", "[DAY 1] Place a sheet of copy paper on a stainless-s...
85,305
Immunostaining of iPSC-derived neurons for quantification of synaptic proteins
4
dx.doi.org/10.17504/protocols.io.8epv5x91ng1b/v1
https://www.protocols.click/view/immunostaining-of-ipsc-derived-neurons-for-quantif-cxizxkf6
Dan Dou, Erika Holzbaur
TITLE: Immunostaining of iPSC-derived neurons for quantification of synaptic proteins AUTHORS: Dan Dou, Erika Holzbaur [DESCRIPTION] Here, we fix, permeabilize, and stain human iPSC-derived neurons for the purpose of observing and quantifying somal proteins of interest. For preceding culture of neurons, see "Protocol:...
["At DIV14, fix human iNeurons in 4% paraformaldehyde supplemented with 4% sucrose for 15 minutes at 37 degrees C", "Wash four times with PBS", "Permeabilize for 15 minutes in 0.2% Triton-X in PBS", "Block for 1 hour with 5% goat serum and 1% BSA in PBS", "Incubate in primary antibody diluted in blocking solution at ro...
49,458
Processing a Viral Metagenome Using iVirus
2
dx.doi.org/10.17504/protocols.io.buisnuee
https://www.protocols.io/view/processing-a-viral-metagenome-using-ivirus-buisnuee
Benjamin Bolduc
TITLE: Processing a Viral Metagenome Using iVirus AUTHORS: Benjamin Bolduc [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>A collection of protocols designed to guide the user in processing a viral metagenome from raw sequence data to assembly, and subsequent analysis. The user uses </span><sp...
[]
60,062
Staphilococcus Aureus Sampling
4
dx.doi.org/10.17504/protocols.io.81wgb6pk1lpk/v6
https://www.protocols.io/view/staphilococcus-aureus-sampling-b6v6re9e
Pol Roca Cugat, Olga Sánchez
TITLE: Staphilococcus Aureus Sampling AUTHORS: Pol Roca Cugat, Olga Sánchez [DESCRIPTION] This protocol is intended to study the affectation of Staphilococcus Aureus, including the MRSA variant. It outlines the basic protocol for a multi-subject study. [GUIDELINES] This protocol is intended to study the affectation...
["[Preparation] Wash your hands with soap. Put on your lab coat, your mask and your goggles or face shield. Make sure your mask is airtight and air cannot escape through the sides.", "[Preparation] Prepare the area where you are going to work. Disinfect the surfaces with the bleach solution.\nThe subjects should not be...
53,732
Immunohistochemical labelling of lower urinary tract afferents in spinal cord
1
dx.doi.org/10.17504/protocols.io.byqcpvsw
https://www.protocols.io/view/immunohistochemical-labelling-of-lower-urinary-tra-byqcpvsw
Janet R Keast, Peregrine B Osborne, John-Paul Fuller-Jackson
TITLE: Immunohistochemical labelling of lower urinary tract afferents in spinal cord AUTHORS: Janet R Keast, Peregrine B Osborne, John-Paul Fuller-Jackson [DESCRIPTION] This protocol is used for immunohistochemical visualisation of cholera toxin subunit B within afferents innervating the lower urinary tract in cryose...
["[Preparation of cryosections] Cryoprotect fixed tissue (L5-S2 spinal cord) in phosphate-buffered saline (PBS; 0.1 M, pH7.2) containing 30% sucrose. This should be performed at 4C, 24-72h prior to cutting.", "[Preparation of cryosections] Embed tissue in cryomold using OCT, freeze in cryostat and cut sections (40 µm),...
76,419
Inoue transformation buffer
4
null
https://www.protocols.io/view/inoue-transformation-buffer-cnvbve2n
Andreas Sagen
TITLE: Inoue transformation buffer AUTHORS: Andreas Sagen [DESCRIPTION] The Inoue method of transformation is a chemical transformation method of prokaryotes (bacteria), which add significant improvements to methods such as the classic Calcium chloride transformation method. [STEPS] SECTION: 10x Inoue transformation ...
["[10x Inoue transformation buffer] Add 80 mL distilled water to a 100 mL tube", "[10x Inoue transformation buffer] Measure and add 10.89 g Manganese chloride, 2.2 g Calcium chloride, 3.0 g PIPES and 18.65 g Potassium chloride\n\nMaterials:", "[10x Inoue transformation buffer] Add distilled water to 100 mL", "[10x Inou...
83,027
Isolation and storage of PBMCs from human peripheral blood samples
1
dx.doi.org/10.17504/protocols.io.14egn3bzml5d/v1
https://www.protocols.click/view/isolation-and-storage-of-pbmcs-from-human-peripher-cvbtw2nn
Yunge Zhao, Joseph Rabin, Joel Linden, Christine L Lau
TITLE: Isolation and storage of PBMCs from human peripheral blood samples AUTHORS: Yunge Zhao, Joseph Rabin, Joel Linden, Christine L Lau [DESCRIPTION] This protocol descried detailed steps on how to isolate and store PBMCs from human peripheral blood samples. By following this protocol, you can isolate and store PBM...
["[Isolation and storage of PBMCs from human peripheral blood samples] Collect 2 tubes of 10ml of blood in EDTA tubes and transport to the laboratory in an ice box.", "[Isolation and storage of PBMCs from human peripheral blood samples] Spin samples at 1600 rpm (2500 x G) for 10 minutes in collection tubes.", "[Isolati...
51,771
Protocol 1: Clinical and demographic information
1
dx.doi.org/10.17504/protocols.io.bws3pegn
https://www.protocols.io/view/protocol-1-clinical-and-demographic-information-bws3pegn
Sophie Adler, Mathilde Ripart, Meld Project, Konrad Wagstyl
TITLE: Protocol 1: Clinical and demographic information AUTHORS: Sophie Adler, Mathilde Ripart, Meld Project, Konrad Wagstyl [DESCRIPTION] The MELD Project is an international collaboration aiming to create open-access, robust and generalisable tools for FCD detection. This MRI data protocol details 1) How to ge...
["[Access to the RedCap Database] Anonymised clinical and demographic information for all participants should be entered by filling in the participant information survey on Redcap.\n\nEach participating site needs to send us the institutional email addresses of the individuals who will be carrying out data entry. Accou...
103,458
RNA Isolation of Human Osteochondral, Subchondral Bone and Cartilage Tissues
0
dx.doi.org/10.17504/protocols.io.6qpvr8m2zlmk/v1
https://www.protocols.io/view/rna-isolation-of-human-osteochondral-subchondral-b-dhaa32ae
Ann-Marie Hageny, Merissa Olmer, Martin Lotz
TITLE: RNA Isolation of Human Osteochondral, Subchondral Bone and Cartilage Tissues AUTHORS: Ann-Marie Hageny, Merissa Olmer, Martin Lotz [DESCRIPTION] This protocol demonstrates how to perform RNA isolation from fresh snapfrozen Human Osteochondral Tissue, Subchondral Bone and Cartilage tissues. [STEPS] SECTION: Ti...
["[Tissue Homogenization] Precool Benchtop Centrifuge and make sure temperature is at 4℃.", "[Tissue Homogenization] Prepare tools, mortar and pestle, and work surface by spraying down with RNAse Away.", "[Tissue Homogenization] Weigh out 110-150mg of fresh or snapfrozen tissue. If you have to cut a smaller piece of sn...
52,279
Detection of Cryptosporidium in stool sample by PCR-RFLP
4
dx.doi.org/10.17504/protocols.io.bxaxpifn
https://www.protocols.io/view/detection-of-cryptosporidium-in-stool-sample-by-pc-bxaxpifn
botchiesenyo
TITLE: Detection of Cryptosporidium in stool sample by PCR-RFLP AUTHORS: botchiesenyo [DESCRIPTION] Nested PCR-RFLP adapted from Nichols, R.A.B., Campbell, B.M. and Smith, H.V., 2003. Identification of Cryptosporidium spp. oocysts in United Kingdom noncarbonated natural mineral waters and drinking waters by using a ...
["[Nested PCR] Extract whole genomic DNA from stool samples. \n\nPositive control DNA for Cryptosporidium parvum may be obtained from ATCC. Alternatively you can extract genomic DNA from wild type Cryptosporidium parvum purchased from Bunchgrass Farms (Deary, Idaho, USA), Waterborne Inc (New Orleans, Louisiana, USA), o...
70,236
Application of PHYTO-PAM-II (Compact Version) on Aureococcus anophagefferens cultures for photosynthetic efficiency and quantum yield of PSII 
1
dx.doi.org/10.17504/protocols.io.yxmvm2785g3p/v1
https://www.protocols.io/view/application-of-phyto-pam-ii-compact-version-on-aur-cgt4twqw
Emily E. Chase, Gwen Stark, Alex Truchon, Steven W Wilhelm
TITLE: Application of PHYTO-PAM-II (Compact Version) on Aureococcus anophagefferens cultures for photosynthetic efficiency and quantum yield of PSII  AUTHORS: Emily E. Chase, Gwen Stark, Alex Truchon, Steven W Wilhelm [DESCRIPTION] A protocol used to acquire photosynthetic efficiency (Fv/Fm) and quantum yield of photo...
["[SAMPLE PREPARATION] Prepare 3 mL of each culture of Aureococcus anophagefferens to be measured in appropriately labelled, separate 15 mL Falcon tubes. Additionally, prepare at least one sample to carry out the auto-gain function if you do not know the appropriate gain (see Step 3.1).", "[SAMPLE PREPARATION] For acqu...
41,768
Protocol to apply saliva samples to quantum mechanical tunneling current real-time point-of-care COVID-19 sensor.
4
dx.doi.org/10.17504/protocols.io.bk2gkybw
https://www.protocols.io/view/protocol-to-apply-saliva-samples-to-quantum-mechan-bk2gkybw
Massood Tabib-Azar
TITLE: Protocol to apply saliva samples to quantum mechanical tunneling current real-time point-of-care COVID-19 sensor. AUTHORS: Massood Tabib-Azar [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is used to operate an electronic rapid (1 min) point-of-care SARS-CoV-2 sensor</div></div...
["Apply a small amount (~1 uL) of saliva on the sensor element at room temperature.", "2. Wait for 30 s. The sensor sytem wil turn on a green LED to alert you at the end of the 30 s.", "Measure the sensor I-V using the paired smart phone APP or click on the \"Read\" botton on the sensor system.", "Apply 60 C water on t...
null
null
null
dx.doi.org/10.17504/protocols.io.ieacbae
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p> </p> <p>The protocol described here is an adaptation from this paper:</p> <p> </p> <p>Sun, Z. Z., Hayes, C. A., Shin, J., Caschera, F., Murray, R. M., Noireaux, V. Protocols for Implementing an <em>Escherichia coli</em> Based TX-TL Cell-Free Expression System for Synthetic B...
[]
100,654
Tissue processing and freezing after surgery
1
null
https://www.protocols.io/view/tissue-processing-and-freezing-after-surgery-dein3cde
Bettina Ergün
TITLE: Tissue processing and freezing after surgery AUTHORS: Bettina Ergün [DESCRIPTION] The aim of this protocol is to document the processing of fresh tissue after surgery. The collected tissue must also be documented in the files which are linked down below The protocol descibes in detail how to process the res...
["[Sample processing in the lab] Fresh tissue should be processed on the same day if possible, but can be stored at 4°C for a maximum of overnight.\nTo reduce pathogens, the tissue should be incubated for 60 min atRoom temperature", "[Sample processing in the lab]", "[Sample processing in the lab]", "[Sample processing...
49,399
Efficient third generation lentiviral particle production
4
dx.doi.org/10.17504/protocols.io.bugxntxn
https://www.protocols.io/view/efficient-third-generation-lentiviral-particle-pro-bugxntxn
Michelle Newbery, Simon Maksour, Amy Hulme, Neville Ng, Mirella Dottori, Lezanne Ooi
TITLE: Efficient third generation lentiviral particle production AUTHORS: Michelle Newbery, Simon Maksour, Amy Hulme, Neville Ng, Mirella Dottori, Lezanne Ooi [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The overexpression of a gene of interest by third generation lent...
["[Lentiviral transfection]\nMaintain HEK293T cells in 0.15 mL/cm2 (e.g. 12 mL in 75 cm2 flask) animal product free FreeStyle 293 Expression Medium, foetal serum free DMEM/F12 + 2 mM GlutaMAX + 5% Knock Out Serum Replacement or DMEM/F12 + 2 mM GlutaMAX + 5% FBS. Include 50 U/mL Penicillin and Streptomycin to reduce ris...
72,237
Protocol 6: Selecting for Spizellomyces punctatus transformants
4
dx.doi.org/10.17504/protocols.io.dm6gpjjdpgzp/v1
https://www.protocols.io/view/protocol-6-selecting-for-spizellomyces-punctatus-t-cismuec6
Sarah M Prostak, Edgar M Medina, Erik Kalinka, Lillian Fritz-Laylin
TITLE: Protocol 6: Selecting for Spizellomyces punctatus transformants AUTHORS: Sarah M Prostak, Edgar M Medina, Erik Kalinka, Lillian Fritz-Laylin [DESCRIPTION] Once the co-culture IM plates have grown for four days, it is time to select Sp transformants. Selecting these transformants requires harvesting the cells fr...
["[Steps] For each plasmid transformed, prepare a 50 mL conical with 30 mL of DS solution.", "[Steps] From the appropriate conical for the plate to be harvested, remove 1 mL of DS.", "[Steps] Distribute the DS among the four quadrants of the corresponding IM plate.", "[Steps] Incubate at Room temperature for 5 min.", "...
73,284
Chloroform-methanol protein precipitation from microalgae and Pierce BCA assay
4
dx.doi.org/10.17504/protocols.io.yxmvm2e25g3p/v1
https://www.protocols.io/view/chloroform-methanol-protein-precipitation-from-mic-cjtcuniw
Ying-Yu Hu, Christopher Lord, Zoe V Finkel
TITLE: Chloroform-methanol protein precipitation from microalgae and Pierce BCA assay AUTHORS: Ying-Yu Hu, Christopher Lord, Zoe V Finkel [DESCRIPTION] Chlorophyll, phospholipids, sucrose, glycerol and some detergent in crude protein extracted from microalgae can interfere the Pierce BCA protein assay. In order to rem...
["[Reagent preparation] Tris buffer 5 mM (pH 8.0)\nAdd 500 µL 1 MpH 8.0 Tris into 100 mL MilliQ water", "[Reagent preparation] 20% Sarcosine\nDilute 2 part 30% N-lauroylsarcosine sodium salt with 1 part 5 mM (pH 8.0) Tris buffer", "[Protein precipitation] Thaw protein extract", "[Protein precipitation] Turn on refr...
23,198
cDNA synthesis using the Applied BiosystemsTM High-Capacity cDNA Reverse Transcription Kit
null
dx.doi.org/10.17504/protocols.io.2v6ge9e
null
Lisa-Maria Rosenthal, Giang Tong, Katharina Schmitt
TITLE: cDNA synthesis using the Applied BiosystemsTM High-Capacity cDNA Reverse Transcription Kit AUTHORS: Lisa-Maria Rosenthal, Giang Tong, Katharina Schmitt [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">cDNA synthesis using the Applied Biosystems</span><span sty...
["Allow the kit components to thaw on ice.Prepare the 2X RT master mix on ice. AB1ComponentVolume (µL)210X RT Buffer2.0325X dNTP Mix (100 mM)0.8410X RT Random Primers2.05MultiScribeTMReverse Transcriptase1.06RNase Inhibitor1.07Nuclease-free H2O3.28Total per reaction10.0 Place the 2X RT master mix on ice and mix gentl...
43,852
Analysis of the Chromosomal Localization of Yeast SMC Complexes by Chromatin Immunoprecipitation
2
dx.doi.org/10.17504/protocols.io.bn3kmgkw
https://www.protocols.io/view/analysis-of-the-chromosomal-localization-of-yeast-bn3kmgkw
Vasso Makrantoni, Daniel Robertson, Adele L. Marston
TITLE: Analysis of the Chromosomal Localization of Yeast SMC Complexes by Chromatin Immunoprecipitation AUTHORS: Vasso Makrantoni, Daniel Robertson, Adele L. Marston [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">A plethora of biological processes like gene transcription, DNA replication, DNA recom...
[]
43,121
Multitissue DNA methylome profiling during onset of salmon maturation
5
dx.doi.org/10.17504/protocols.io.bncrmav6
https://www.protocols.io/view/multitissue-dna-methylome-profiling-during-onset-o-bncrmav6
Amin Mohamed, Bradley Evans, Antonio Reverter, James Kijas
TITLE: Multitissue DNA methylome profiling during onset of salmon maturation AUTHORS: Amin Mohamed, Bradley Evans, Antonio Reverter, James Kijas [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Background:</div><div class = "text-block">Atlantic salmon farming promotes growth in conditions which mean...
["[Genomic DNA isolation, WGBS library preparation and sequencing]\nTissue samples were snap frozen in liquid Nitrogen and stored at −80 °C until genomic DNA (gDNA) was extracted using DNeasy blood and tissue kit (QIAGEN). Tissues were lysed in 360 µL of lysis solution on a Precellys 24 homogenizer for 30s at 4.0 ms−1....
68,221
Laser Capture Microdissection of Tissue Functional Units for microPOTS Top-Down Proteomics
4
dx.doi.org/10.17504/protocols.io.ewov1nrz7gr2/v1
https://www.protocols.io/view/laser-capture-microdissection-of-tissue-functional-ceu5tey6
James M Fulcher, Yen-Chen Liao
TITLE: Laser Capture Microdissection of Tissue Functional Units for microPOTS Top-Down Proteomics AUTHORS: James M Fulcher, Yen-Chen Liao [DESCRIPTION] In this protocol, we describe a detailed procedure for high-resolution top-down proteomic analysis of human pancreatic and kidney tissue sections using a spatial prote...
["[Tissue collection] Load DMSO as capturing media onto the microPOTS chip. Microdroplet processing in one pot for trace samples (microPOTS) chips are polyproplyene chips designed for benchtop manipulation of low sample volumes ( < 5 μL).", "[Tissue collection] Pipette 1 µL of DMSO onto each microPOT chip well", "[Tis...