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null
null
null
dx.doi.org/10.17504/protocols.io.chmt45
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This is the protocol for the Q5® Site-Directed Mutagenesis Kit without competent cells [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
31,074
Chloral Hydrate Seed Clearing
null
dx.doi.org/10.17504/protocols.io.bakaicse
null
Gabrielle Sandstedt, Andrea Sweigart
TITLE: Chloral Hydrate Seed Clearing AUTHORS: Gabrielle Sandstedt, Andrea Sweigart [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">To characterize early seed development in Mimulus (1-5 days after pollination), we clear seeds with chloral hydrate and quickly obtain images of embryo and endosperm dev...
["[Dissecting fruit and clearing]\nEmasculate a bud from some maternal plant. 2 to 3 days later pollinate by selfing/outcrossing or use an unfertilized fruit.", "[Dissecting fruit and clearing]\nRemove the developing fruit 1 to 5 days after pollination or 2 to 3 days after emasculation. In Mimulus, this protocol is use...
58,684
Detailed Western Blotting (Immunoblotting) Protocol
4
dx.doi.org/10.17504/protocols.io.b5i4q4gw
https://www.protocols.io/view/detailed-western-blotting-immunoblotting-protocol-b5i4q4gw
Rasheed Sule, Gabriela Rivera, Aldrin V Gomes
TITLE: Detailed Western Blotting (Immunoblotting) Protocol AUTHORS: Rasheed Sule, Gabriela Rivera, Aldrin V Gomes [DESCRIPTION] Western Blotting, which is probably better referred to as immunoblotting, is one of the most commonly used biological methods worldwide. This technique is capable of detecting an indivi...
["[Sample preparation] Sample preparation can be simple or complex, depending on the source of the sample and the location of the target protein. It is recommended that you consult a dedicated article with procedures for optimal sample preparation.\nThis protocol assumes you have already prepared your sample and are re...
67,448
Via Keto Apple Gummies Australia Reviews- Price or Chemist Warehouse Buy
1
dx.doi.org/10.17504/protocols.io.bp2l61qndvqe/v1
https://www.protocols.io/view/via-keto-apple-gummies-australia-reviews-price-or-cd4ys8xw
health
TITLE: Via Keto Apple Gummies Australia Reviews- Price or Chemist Warehouse Buy AUTHORS: health [DESCRIPTION] Via Keto Apple Gummies - Natural products are always a better choice due to their medicinal potential and guaranteed safety. [STEPS] 1. Via Keto Apple Gummies is a weight loss component primarily based on a...
["Via Keto Apple Gummies is a weight loss component primarily based on a ketogenic food regimen that includes essential nutrients together with omega-3s, multivitamins, as well as protein to offer speedy weight management effects without facet effects. Official Web: https://www.mynewsdesk.com/nealthnewscart/pressreleas...
35,720
SNARE-seq2
1
dx.doi.org/10.17504/protocols.io.be5gjg3w
https://www.protocols.io/view/snare-seq2-be5gjg3w
Nongluk Plongthongkum, Dinh H Diep, Song Chen, Blue Lake, Kun Zhang
TITLE: SNARE-seq2 AUTHORS: Nongluk Plongthongkum, Dinh H Diep, Song Chen, Blue Lake, Kun Zhang [DESCRIPTION] To study the heterogeneity of complex tissues by joint profiling of gene expression and its regulation, we require an accurate and high-throughput method. Here we described improved high-throughput combinato...
["[Reagent setup] 40% (wt/vol) PEG 6000. Weigh 16.0 g of PEG 6000 in 50 mL tube. Add nuclease-free water and bring the total volume to 40 mL. Rotate the tube at room temperature until PEG 6000 completely dissolved. Spin down the tube at 200 g for 2 min, at room temperature to remove the tiny bubble. CRITICAL: 40% (wt/v...
27,390
Tissue Procurement: Biosafety Guidelines
1
dx.doi.org/10.17504/protocols.io.6y6hfze
https://www.protocols.io/view/tissue-procurement-biosafety-guidelines-6y6hfze
Kerry Wiles
TITLE: Tissue Procurement: Biosafety Guidelines AUTHORS: Kerry Wiles [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>The biosafety guidelines in this protocol are based on the Center for Disease Control's </span><span style = "font-style:italic;">Biosafety in Microbiology and Biomedical Labora...
["[Tissue Procurement: Biosafety Guidelines]\nCollection is done by trained personnel and all products are handled separately and under as sterile conditions as possible. You will always take the necessary precautions for your safety, BUT also take the necessary precautions to protect the tissue FROM you.", "[Tissue Pr...
41,996
CollectingCitationsfromText
5
dx.doi.org/10.17504/protocols.io.bk9kkz4w
https://www.protocols.io/view/collectingcitationsfromtext-bk9kkz4w
Rebecca Hedreen
TITLE: CollectingCitationsfromText AUTHORS: Rebecca Hedreen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Basic steps and scripts used for translating text citations to bibtex files suitable for loading into citation management software or citation analysis scripts. 3 publically available webpage ...
["[Preparing text file]\nCopy citations from the source document(s) into a text (.txt) document.", "[Preparing text file]\nEdit the text document so that each citation is on a separate line with one blank line between each citation. Not all the scripts require a blank line between citations, but it does improve readabi...
10,256
1% Agarose Gel Electrophoresis Prep
null
dx.doi.org/10.17504/protocols.io.m9qc95w
null
Claire Rycroft, Ben JG. Sutherland
TITLE: 1% Agarose Gel Electrophoresis Prep AUTHORS: Claire Rycroft, Ben JG. Sutherland [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Ararose (1%) gel electrophoresis for genomic DNA quality checking.</div><div class = "text-block">This protocol comes with no guarantees from the authors.</div></div...
["[Gel Prep]\nWeigh out 2.5g of powdered agarose.", "[Gel Prep]\nAdd 250mL of 1XTAE buffer to a 1.0L Erlenmeyer flask.Note:Be mindful to use best practices when making solutions. Make sure the glassware you are using has been properly cleaned and has been pre-rinsed with whatever solution you will be working with, in t...
null
null
null
dx.doi.org/10.17504/protocols.io.magc2bw
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
70,154
fastANI analysis protocol
5
null
https://www.protocols.io/view/fastani-analysis-protocol-cgritv4e
Jamie Harrison, David J Studholme
TITLE: fastANI analysis protocol AUTHORS: Jamie Harrison, David J Studholme [DESCRIPTION] this is the protocol to conduct ani analysis between groups of genomes using fastANI and produce the heatmap figure in R using the pheatmap package [STEPS] SECTION: create directories 1.2. mkdir analysis query_genomes reference_...
["[create directories] mkdir analysis query_genomes reference_genomes", "[create directories] move to query dir", "[create directories]", "[create directories]", "[create directories]", "Create directory structure and link files", "[create directories] move to reference dir", "[create directories]", "[create directorie...
22,753
Microbiological contamination of young children’s hands in rural Bangladesh: Associations with child age and observed hand cleanliness as proxy
null
dx.doi.org/10.17504/protocols.io.2f9gbr6
null
Sarker Masud Parvez, Rashidul Azad, Amy J. Pickering, Laura H Kwong, Benjamin F. Arnold, Musarrat Jabeen Rahman, Md. Zahidur Rahman, Mahfuja Alam, Debashis Sen, Sharmin Islam, Mahbubur Rahman, John M. Colford, Jr, Stephen P. Luby, Leanne Unicomb, Ayse Ercumen
TITLE: Microbiological contamination of young children’s hands in rural Bangladesh: Associations with child age and observed hand cleanliness as proxy AUTHORS: Sarker Masud Parvez, Rashidul Azad, Amy J. Pickering, Laura H Kwong, Benjamin F. Arnold, Musarrat Jabeen Rahman, Md. Zahidur Rahman, Mahfuja Alam, Debashis Sen,...
[]
44,408
Fluorescence Recovery after Photobleaching (FRAP)  in adult C. elegans nuclei
4
dx.doi.org/10.17504/protocols.io.bpkymkxw
https://www.protocols.io/view/fluorescence-recovery-after-photobleaching-frap-in-bpkymkxw
Laura Breimann, Kustrim Cerimi, Vic-Fabienne Schumann, Stephan Preibisch, Sevinc Ercan, Andrew Woehler
TITLE: Fluorescence Recovery after Photobleaching (FRAP)  in adult C. elegans nuclei AUTHORS: Laura Breimann, Kustrim Cerimi, Vic-Fabienne Schumann, Stephan Preibisch, Sevinc Ercan, Andrew Woehler [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Protocol for Fluorescence Recovery after Photoble...
["[Preparation of agarose pads]\nPrepare two glass slides as spacers by taping one layer of lab tape along the slides' long side. Tape them to the table, separated to leave space for one glass slide in between them.", "[Preparation of agarose pads]\nDissolve agarose in M9 to create solution and slowly melt in microwav...
74,741
Recombinant expression and purification of HIV-1 RT
4
dx.doi.org/10.17504/protocols.io.3byl4jzyjlo5/v1
https://www.protocols.io/view/recombinant-expression-and-purification-of-hiv-1-r-ck8vuzw6
Javiera A Avilés, Tamara Matute, Isaac Núñez, Maira Rivera, Javiera Reyes, Jenny Molloy, Cesar A Ramirez-Sarmiento, Fernan Federici
TITLE: Recombinant expression and purification of HIV-1 RT AUTHORS: Javiera A Avilés, Tamara Matute, Isaac Núñez, Maira Rivera, Javiera Reyes, Jenny Molloy, Cesar A Ramirez-Sarmiento, Fernan Federici [DESCRIPTION] This protocol has been optimized for the recombinant expression of HIV-1 RT. The sequence of the plasmid...
["[DAY 1 – Plasmid transformation] Transform 100 ng of plasmid containing HIV-1 RT into E. coli BL21(DE3) competent cells using either heat shock or electroporation.", "[DAY 1 – Plasmid transformation] Spread transformed cells in LB Agar plates supplemented with 0.05 mg/mL Kan. Grow plate overnight at 37 °C.", "[DAY 2 ...
68,477
Modified Qiagen DNeasy Blood and Tissue extraction method (Cat. No. / ID: 69506) for eDNA extraction from filters (Nitrocellulose Mixed Ester membrane filters or similar)
4
dx.doi.org/10.17504/protocols.io.8epv59y4jg1b/v1
https://www.protocols.io/view/modified-qiagen-dneasy-blood-and-tissue-extraction-ce45tgy6
Yoamel Milián-García
TITLE: Modified Qiagen DNeasy Blood and Tissue extraction method (Cat. No. / ID: 69506) for eDNA extraction from filters (Nitrocellulose Mixed Ester membrane filters or similar) AUTHORS: Yoamel Milián-García [DESCRIPTION] The commercial Qiagen DNeasy Blood and Tissue kit constitutes one of the most used kits in Molecu...
["Allow a filter to thaw on a sterile Petri Dish (e.g., Fisher Scientific Catalog No. FB0875713 or FB0875713A) and cut into quarters (halves can be used to decrease the number of extractions per filter). Cut the filter quarters (or halves) into strips using sterile razor blades. Filter strip manipulations at this step ...
null
null
null
dx.doi.org/10.17504/protocols.io.qywdxxe
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Protocol for collecting gas for measurement of carbon isotope descrimination during photosynthesis.</p> <p> </p> <p>See the following references for more background:</p> <p>H. Griffiths (1993) <a href="https://link.springer.com/book/10.1007/978-94-011-1566-7" target="_blank" ...
[]
30,987
CIDC_S16_LC_MS_Celegans_Extraction_Protocol
1
dx.doi.org/10.17504/protocols.io.bahjib4n
https://www.protocols.io/view/cidc-s16-lc-ms-celegans-extraction-protocol-bahjib4n
Brianna M Garcia, Carter Asef
TITLE: CIDC_S16_LC_MS_Celegans_Extraction_Protocol AUTHORS: Brianna M Garcia, Carter Asef [DESCRIPTION] A sample preparation protocol for lyophlized C. elegans samples to be analyzed via LC-MSMS [STEPS] SECTION: Homogenization 1. Samples are removed from -80 °C SECTION: Homogenization 2. (3) 2.0mm zirconium oxide b...
["[Homogenization] Samples are removed from -80 °C", "[Homogenization] (3) 2.0mm zirconium oxide beads and ~ 75 µL volume of 0.5mm glass beads are added to each sample tube.", "[Homogenization] Samples are placed in Tissuelyser II using adapter trays chilled at -80C and homogenized at 1800rpm for 3 min.", "[Homogeniz...
36,237
Footprint-Free Genome Editing of iPSC Using Alt-R CRISPR/Cas9
null
dx.doi.org/10.17504/protocols.io.bfmmjk46
https://www.protocols.io/view/footprint-free-genome-editing-of-ipsc-using-alt-r-bfmmjk46
Jacob Marsh, Rj Martinez, Celeste Karch
TITLE: Footprint-Free Genome Editing of iPSC Using Alt-R CRISPR/Cas9 AUTHORS: Jacob Marsh, Rj Martinez, Celeste Karch [STEPS] ?. [Preparing iPSCs for Nucleofection] Coat 1 well of a 6-well plate with of Matrigel for at 1 ml 37 °C ?. [Preparing iPSCs for Nucleofection] Add of DMEM/F12 to a 15ml conical tue 5 ml ?....
["[Preparing iPSCs for Nucleofection]\nCoat 1 well of a 6-well plate with of Matrigel for at\n1 ml\n37 °C", "[Preparing iPSCs for Nucleofection]\nAdd of DMEM/F12 to a 15ml conical tue\n5 ml", "[Preparing iPSCs for Nucleofection]\nThaw vial of cells in water bath for approximately\n37 °C", "[Preparing iPSCs for Nu...
55,922
My demo protocol
1
null
https://www.protocols.io/view/my-demo-protocol-b2usqewe
Gerard Weatherby
TITLE: My demo protocol AUTHORS: Gerard Weatherby [DESCRIPTION] API test [STEPS] SECTION: Section 1 1. Do something. SECTION: Section 1 1.1. Start here SECTION: Section 1 1.2.
["[Section 1] Do something.", "[Section 1] Start here", "[Section 1]"]
35,596
Artificial Cerebrospinal Fluid I (ACSF.I)
null
dx.doi.org/10.17504/protocols.io.bezkjf4w
null
Allen Institute for Brain Science
TITLE: Artificial Cerebrospinal Fluid I (ACSF.I) AUTHORS: Allen Institute for Brain Science [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is used to prepare Artificial Cerebrospinal Fluid I (ACSF.I). ACSF.I is used for applications including transcardial perfusion prior to fresh mous...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.gusbwwe
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <div class="page" title="Page 2"> <div class="layoutArea"> <div class="column"> <p>Western blot detection of proteins requires separation of protein mixtures by electrophoresis, followed by transfer of the separated proteins to nitrocellulose or PVDF membranes for detection.The ...
[]
46,008
Human Tissue Slicing and Dissections for Nuclear Isolations
1
dx.doi.org/10.17504/protocols.io.bq6ymzfw
https://www.protocols.io/view/human-tissue-slicing-and-dissections-for-nuclear-i-bq6ymzfw
Allen Institute for Brain Science
TITLE: Human Tissue Slicing and Dissections for Nuclear Isolations AUTHORS: Allen Institute for Brain Science [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol describes the slicing and microdissection procedure on frozen post-mortem human brain tissue to be used to isolate nuclei for su...
[]
93,199
Stereotaxic viral injections and array implantation
1
dx.doi.org/10.17504/protocols.io.14egn31q6l5d/v1
https://www.protocols.io/view/stereotaxic-viral-injections-and-array-implantatio-c69pzh5n
Mai-Anh Vu, mwhowe
TITLE: Stereotaxic viral injections and array implantation AUTHORS: Mai-Anh Vu, mwhowe [DESCRIPTION] We have developed a new micro-fiber array approach capable of chronically measuring and optogenetically manipulating local dynamics across over 100 targeted locations simultaneously in head-fixed and freely moving mice...
["[viral injection] Mice were anesthetized under isoflurane (1-3%) and placed in a stereotaxic frame.", "[viral injection] Mice were injected with AAVs to express genetically encoded proteins for optical measurements and manipulations. Virus (~200-800nL per site) was injected stereotaxically through a pulled glass pipe...
75,970
PCR-NGS for RNA viruses
4
dx.doi.org/10.17504/protocols.io.x54v9d361g3e/v1
https://www.protocols.io/view/pcr-ngs-for-rna-viruses-cnfavbie
Masayasu Misu, Tomoki Yoshikawa, Satoko Sugimoto, Yuki Takamatsu, Takeshi Kurosu, Yukiteru Ouji, Masahide Yoshikawa, Masayuki Shimojima, Hideki Ebihara, Masayuki Saijo
TITLE: PCR-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 PCR-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 RNA extraction] The viral genomic RNA extraction is performed usin...
100,136
Single-cell single-unit recordings in vitro
0
dx.doi.org/10.17504/protocols.io.yxmvme269g3p/v1
https://www.protocols.io/view/single-cell-single-unit-recordings-in-vitro-dd2g28bw
Nicola Biagio Mercuri
TITLE: Single-cell single-unit recordings in vitro AUTHORS: Nicola Biagio Mercuri [DESCRIPTION] Single units in slices [STEPS] SECTION: Single-cell single-unit recordings 1. The slice is placed in a submerged recording chamber in continuously flowing ACSF (34°C) saturated with O2/CO2 gas mixture (95/5%). SECTION: Sin...
["[Single-cell single-unit recordings] The slice is placed in a submerged recording chamber in continuously flowing ACSF (34°C) saturated with O2/CO2 gas mixture (95/5%).", "[Single-cell single-unit recordings] ACSF composition (in mM): 126 NaCl, 2.5 KCl,1.2 MgCl2, 1.2 NaH2PO4, 2.4 CaCl2, 10 glucose and 25 NaHCO3.", "[...
null
null
null
dx.doi.org/10.17504/protocols.io.hjsb4ne
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] A protocol to separate chloroplasts from diatom cells using ammonium fluoride to permeate the silica frustrule and a percoll gradient to separate the plastid from other cellular components. [BEFORE_START] Make sure all buffers are prepared according to the instructions in "Buff...
[]
18,288
SYSB 3036 W11: Gene Expression and RNA-Seq
null
dx.doi.org/10.17504/protocols.io.v4qe8vw
null
Frank Aylward
TITLE: SYSB 3036 W11: Gene Expression and RNA-Seq AUTHORS: Frank Aylward [STEPS] ?. [Getting started] Last tutorial we went over basic read mapping protocols using bowtie and SAMtools. We used data from a infection experiment that used Mycobacterium smegmatis and a bacteriophage called D29. We examined only one time-p...
["[Getting started]\nLast tutorial we went over basic read mapping protocols using bowtie and SAMtools. We used data from a infection experiment that used Mycobacterium smegmatis and a bacteriophage called D29. We examined only one time-point that took place 15 minutes after infection. Today we will finish analyzing th...
null
null
null
dx.doi.org/10.17504/protocols.io.gzebx3e
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Gibson assembly using NEBuilder HiFi DNA assembly kit. Specification of the original NEB protocol.</p> [STEPS] ?. ?. ?. ?.
[]
98,973
Brain Data Alchemy Project: Meta-Analysis of Re-Analyzed Public Transcriptional Profiling Data in the Gemma Database
0
dx.doi.org/10.17504/protocols.io.j8nlk84jxl5r/v1
https://www.protocols.io/view/brain-data-alchemy-project-meta-analysis-of-re-ana-dcv52w86
Megan Hagenauer, Cosette Rhoads, Jinglin Xiong, Duy Manh Nguyen, Erin Hernandez, Annaka Saffron, Amrita Kondur, Elizabeth Flandreau
TITLE: Brain Data Alchemy Project: Meta-Analysis of Re-Analyzed Public Transcriptional Profiling Data in the Gemma Database AUTHORS: Megan Hagenauer, Cosette Rhoads, Jinglin Xiong, Duy Manh Nguyen, Erin Hernandez, Annaka Saffron, Amrita Kondur, Elizabeth Flandreau [DESCRIPTION] Over the past two decades, transcription...
["[Project Preparation: Environment Set-Up] Install R and RStudio", "[Project Preparation: Environment Set-Up] Install R", "[Project Preparation: Environment Set-Up] Install R Studio", "[Project Preparation: Environment Set-Up] Follow the Brain Data Alchemy code repository on Github", "[Project Preparation: Environment...
94,402
Chemogenetic modulation of catecholaminergic neurons
1
dx.doi.org/10.17504/protocols.io.rm7vzxdz5gx1/v1
https://www.protocols.io/view/chemogenetic-modulation-of-catecholaminergic-neuro-c8faztie
Cristian González-Cabrera, Csilla Novák, Maria P. Contreras, Ernesto Durán, Matthias Prigge
TITLE: Chemogenetic modulation of catecholaminergic neurons AUTHORS: Cristian González-Cabrera, Csilla Novák, Maria P. Contreras, Ernesto Durán, Matthias Prigge [DESCRIPTION] This protocol outlines a method for modulating noradrenergic and dopaminergic neuronal activity in the LC and SNc-VTA, respectively, using Cre-d...
["[Stereotactic injection] Stereotactic injection of viruses into DbH-Cre (JAX# 033953) or DAT-Cre (JAX# 006660) animals\n\nViral Construct:\npAAV-hSyn-DIO-hM4D(Gi)-mCherry (Addgene link) \nVolume: 400ul\npAAV-hSyn-DIO-hM3D(Gq)-mCherry (Addgene link)\nVolume: 400ul\n\n(in our experiment we mix our DIO-hTyr Virus 1:1 w...
null
null
null
dx.doi.org/10.17504/protocols.io.kb6csre
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?.
[]
52,924
MicroCT protocols for scanning of embryos and juvenile Hexaplex trunculus
4
dx.doi.org/10.17504/protocols.io.bxw4ppgw
https://www.protocols.io/view/microct-protocols-for-scanning-of-embryos-and-juve-bxw4ppgw
Eva Chatzinikolaou, Kleoniki Keklikoglou
TITLE: MicroCT protocols for scanning of embryos and juvenile Hexaplex trunculus AUTHORS: Eva Chatzinikolaou, Kleoniki Keklikoglou [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Micro-computed tomography (micro-CT) is a high-resolution 3D-imaging technique which is now increasingly applied i...
["[Sample preparation]\nAnesthetization of embryos and juvenile gastropods with 7% MgCl2.", "[Sample preparation]\nSample placement inside a plastic white pipette tip without any scanning medium (in the air).", "[microCT scanning]", "[microCT scanning]\nScanning parameters for embryos and juvenile Hexaplex trunculusVol...
25,105
IVF Bioscience Bovine Slaughterhouse Protocol
null
dx.doi.org/10.17504/protocols.io.4rrgv56
null
Jake Silcock
TITLE: IVF Bioscience Bovine Slaughterhouse Protocol AUTHORS: Jake Silcock [STEPS] ?. [Aspiration and IVM] Dish and Media Preparation ?. [Aspiration and IVM] Add of BO-IVM medium per well to 4-well plates without an Oil overlay and place in the incubator 500 µl ?. [Aspiration and IVM] Additionally, prepare one 35 mil...
["[Aspiration and IVM]\nDish and Media Preparation", "[Aspiration and IVM]\nAdd of BO-IVM medium per well to 4-well plates without an Oil overlay and place in the incubator\n500 µl", "[Aspiration and IVM]\nAdditionally, prepare one 35 millimetre dish with of BO-IVM per 4-well plate and place in the incubator without ...
89,942
Vesicle Fusion on SiO2 Substrates
1
null
https://www.protocols.io/view/vesicle-fusion-on-sio2-substrates-c33wyqpe
Paul Stevenson, Nicole Voce
TITLE: Vesicle Fusion on SiO2 Substrates AUTHORS: Paul Stevenson, Nicole Voce [DESCRIPTION] This protocol outlines the steps to produce large area, uniform supported lipid bilayers on SiO2 substrates with or without patterned features. [STEPS] SECTION: Substrate cleaning 1. Clean plain SiO2 or TiO2-patterned substrat...
["[Substrate cleaning] Clean plain SiO2 or TiO2-patterned substrates by sonicating them in a 1:1 IPA:Acetone mixture for ~2 minutes\nDry with a steady stream of N2", "[Substrate cleaning] Oxygen plasma clean the substrates for 2 minutes to remove any residual surface contamination", "[Vesicle prep] Prepare a 1.2 mg/mL ...
78,250
CellProfiler Pipeline to Obtain Pearson&#39;s correlation coefficients for TMEM55B or pRab10 and RILPL1
5
dx.doi.org/10.17504/protocols.io.rm7vzbqp5vx1/v1
https://www.protocols.io/view/cellprofiler-pipeline-to-obtain-pearson-39-s-corre-cqnivvce
Chloe A Hecht, Suzanne Pfeffer
TITLE: CellProfiler Pipeline to Obtain Pearson&#39;s correlation coefficients for TMEM55B or pRab10 and RILPL1 AUTHORS: Chloe A Hecht, Suzanne Pfeffer [DESCRIPTION] We present here a CellProfiler software pipeline to quantify the intensity of endogenous TMEM55B or pRab10 in LRRK2 R1441C or VPS35 D620N MEF cells transf...
["[Expansion Microscopy TMEM55B and Myc-RILPL1 Correlation] The method involves the following steps:\nStep 1 - Import data and extract metadata from file names\nStep 2 - Group individual channels from each image \nStep 3 - Measure colocalization\nStep 4 - Export the data", "[Expansion Microscopy TMEM55B and Myc-RILPL1 ...
91,023
Sanger Tree of Life Sample Homogenisation: Covaris cryoPREP® Automated Dry Pulverizer
4
dx.doi.org/10.17504/protocols.io.eq2lyjp5qlx9/v2
https://www.protocols.io/view/sanger-tree-of-life-sample-homogenisation-covaris-c45pyy5n
Juan Pablo Narváez-Gómez, Haddijatou Mbye, graeme oatley, Michelle Strickland, Naomi Park, Caroline Howard
TITLE: Sanger Tree of Life Sample Homogenisation: Covaris cryoPREP® Automated Dry Pulverizer AUTHORS: Juan Pablo Narváez-Gómez, Haddijatou Mbye, graeme oatley, Michelle Strickland, Naomi Park, Caroline Howard [DESCRIPTION] This protocol describes the procedure for cryogenic homogenisation of tissue samples using the ...
["[Disruption] Power on the cryoPREP instrument. Ensure all doors to the room are closed and all room occupants are wearing ear defenders. Set the setting dial to “1”.", "[Transfer sample to TissueTUBE] Place the sample into the tissueTUBE TT1 and seal using the adapter and attaching the sample tube on the top.", "[Tra...
43,590
Cell Culture and UV Cross-Linking
4
dx.doi.org/10.17504/protocols.io.bntemeje
https://www.protocols.io/view/cell-culture-and-uv-cross-linking-bntemeje
Clémentine Delan-Forino, David Tollervey
TITLE: Cell Culture and UV Cross-Linking AUTHORS: Clémentine Delan-Forino, David Tollervey [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The RNA exosome complex functions in both the accurate processing and rapid degradation of many classes of RNA in eukaryotes and Archaea. Functional and structur...
["[Growth and Cross-Linking of RNA–Exosome Subunit-His-TEV-ProteinA (HTP) Complexes]\nStreak out HTP-tagged and negative control strains from glycerol stocks onto YPD plates (or SD medium—leucine when required for plasmid maintenance) and incubate at for –.\n30 °C", "[Growth and Cross-Linking of RNA–Exosome Subunit-Hi...
78,340
In vitro GCase activity assay (total cell lysate)
1
dx.doi.org/10.17504/protocols.io.5qpvordxbv4o/v1
https://www.protocols.io/view/in-vitro-gcase-activity-assay-total-cell-lysate-cqrcvv2w
Federico Bertoli, Michela Deleidi
TITLE: In vitro GCase activity assay (total cell lysate) AUTHORS: Federico Bertoli, Michela Deleidi [DESCRIPTION] Glucocerebrosidase is a lysosomal enzyme that catalyzes the hydrolysis of glucosylceramide (GlcCer), a membrane glyco-sphingolipid, to ceramide and glucose. This assay detects GBA activity by using a flu...
["[Sample Lysis] Suspend samples in 50 µL of 1% Triton extraction buffer.", "[Sample Lysis] Homogenize with a Dounce homogenizer for 25 strokes.", "[Sample Lysis] Rotate samples for 30 min at 4 °C.", "[Sample Lysis] Centrifuge at 13500 x g, 4 °C for 15 min.", "[Sample Lysis] Collect supernatants.", "[Substrate prepara...
null
null
null
dx.doi.org/10.17504/protocols.io.dkr4v5
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This lysis buffer is strong enough for lysis most cell lines and wild enough to keep protein interactions. It has been tested on mouse embryonic stem cells, cancer cell lines and overexpressed cells.&nbsp;<br /><br /> [GUIDELINES] <table style="height: 274px;" width="644"> <tbo...
[]
51,745
Preparation of Single Cell Suspension from Human Lung Tissue
1
dx.doi.org/10.17504/protocols.io.bwr9pd96
https://www.protocols.io/view/preparation-of-single-cell-suspension-from-human-l-bwr9pd96
Steven B. Wells, Peter A. Szabo, Basak Ural, Maya M.L. Poon
TITLE: Preparation of Single Cell Suspension from Human Lung Tissue AUTHORS: Steven B. Wells, Peter A. Szabo, Basak Ural, Maya M.L. Poon [DESCRIPTION] This protocol describes a method for the isolation of the immune cells, structural and epithelial cells, and progenitors from human lung sections of about two grams. By...
["[Preparing Mediums and Buffers] Create the following IMDM-FBS-PSQ Media in a 500 mL bottle of IMDM by using the table below: \n Component Volume (mL) Starting Conc.  Final Conc.* IMDM 500 - - Penicillin-Streptomycin-Glutamine 5 100X 1X FBS 50 100% 10%", "[Preparing Mediums and Buffers] Creat...
57,654
HMW DNA extraction for amphipods
4
dx.doi.org/10.17504/protocols.io.b4iwqufe
https://www.protocols.io/view/hmw-dna-extraction-for-amphipods-b4iwqufe
Benoît Vacherie, Karine Labadie
TITLE: HMW DNA extraction for amphipods AUTHORS: Benoît Vacherie, Karine Labadie [DESCRIPTION] Protocol adapted from Qiagen's genomic DNA handbook protocol for the HMW extraction of live or flash-freezed amphipods. [GUIDELINES] To preserve large DNA sizes, never use Vortex and only use Wide-Bore tips. [STEPS] SEC...
["[Tissue homogenization] Prepare the lysis buffer by adding 3 µL of RNase A (100 mg/mL) to 1.5 ml of Buffer G2 per sample.", "[Tissue homogenization] place live or flash-freezed amphipod in 2ml douncer and add 1 ml of lysis buffer", "[Tissue homogenization] gently up and down 10 times with the piston", "[Tissue homog...
null
null
null
dx.doi.org/10.17504/protocols.io.fgtbjwn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol is adapted from the Invitrogen Life Technologies Trizol manual. </p> [BEFORE_START] <ol> <li>Prepare an <strong>RNase-free</strong> working area, wipe down barrels of micropipettes, use <strong>filter tips</strong> and RNase-free microcentrifuge tubes, and alwa...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.n3ddgi6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><em>The MELD Project is an international collaboration aiming to create open-access, robust and generalisable tools for FCD detection. To this end, we will train a neural network classifier on MRI features from FCD patients from multiple centres worldwide.</em></p> <p><strong...
[]
35,038
Lightsheet Tissue Intake - Photodocumentation and Tracking
null
dx.doi.org/10.17504/protocols.io.bef6jbre
https://www.protocols.io/view/lightsheet-tissue-intake-photodocumentation-and-tr-bef6jbre
Seth Currlin, Marda Jorgensen
TITLE: Lightsheet Tissue Intake - Photodocumentation and Tracking AUTHORS: Seth Currlin, Marda Jorgensen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>In order to track tissue position and orientation each sample is photographed when received for lightsheet microscopy (Figure 1, A). Large sa...
["Photograph whole tissue; capture from various angles if possible. A dissection ruler or grid should accompany each photograph.", "Using a razor blade and forceps slice the tissue into sections approximately 2 mm wide. Try to slice in a continuous motion, while applying a gentle downward force.", "Set aside tissue sec...
70,110
Low Biomass, high contamination Illumina DNA prep using DNeasy PowerSoil (Pro) Kit
4
dx.doi.org/10.17504/protocols.io.6qpvr4ey3gmk/v1
https://www.protocols.io/view/low-biomass-high-contamination-illumina-dna-prep-u-cgp6tvre
Tania Kurbessoian, Jason E Stajich, Sonia L. Ghose
TITLE: Low Biomass, high contamination Illumina DNA prep using DNeasy PowerSoil (Pro) Kit AUTHORS: Tania Kurbessoian, Jason E Stajich, Sonia L. Ghose [DESCRIPTION] This is an addendum to the already optimized DNeasy PowerSoil Kit. This can also be applied to the PowerSoil Pro Kit. Use this protocol for Low Biomass or ...
["Add sample into the PowerBead tube.", "Add 60μL of Solution C1.", "Make sure the sample has not precipitated. If so, heat sample to 60ºC until precipitate is dissolved into solution.", "Vortex to mix and incubate at 65ºC for 10 minutes.", "Bead beat at \"homogenize\" setting: 90 seconds bead beating, 60 seconds rest,...
63,998
MEMBER_OWNER 002
1
null
https://www.protocols.io/view/member-owner-002-caq6sdze
rober
TITLE: MEMBER_OWNER 002 AUTHORS: rober [DESCRIPTION] Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque hendrerit mauris quis nibh tincidunt pellentesque. Praesent congue justo vitae molestie dictum. Vivamus odio ipsum, consectetur eu nisi nec, posuere interdum metus. Class aptent taciti sociosqu ad li...
["Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque hendrerit mauris quis nibh tincidunt pellentesque. Praesent congue justo vitae molestie dictum. Vivamus odio ipsum, consectetur eu nisi nec, posuere interdum metus. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenae...
87,307
Sanger Tree of Life Fragmented DNA clean up: Automated SPRI
4
dx.doi.org/10.17504/protocols.io.q26g7p1wkgwz/v1
https://www.protocols.io/view/sanger-tree-of-life-fragmented-dna-clean-up-automa-czhjx34n
graeme oatley, Filipa Sampaio, Caroline Howard
TITLE: Sanger Tree of Life Fragmented DNA clean up: Automated SPRI AUTHORS: graeme oatley, Filipa Sampaio, Caroline Howard [DESCRIPTION] This protocol describes the automated clean up and shorter fragment removal from fragmented DNA following the Sanger Tree of Life HMW DNA Fragmentation protocols, using PacBio AMPure...
["[Laboratory protocol] Normalise the volumes of all sheared DNA samples to the sample with the largest volume.", "[Laboratory protocol] Set-up the KingFisher™ plates for the automated SPRI as detailed below:\n\n Plate namePlate typeReagent(s) requiredTip plateKingFisher‱ Apex 1 mL 96-well deep-well plateKingFisher‱ ...
93,933
Aggregation propensity of α-synuclein seeds in primary neurons
4
dx.doi.org/10.17504/protocols.io.81wgbxe13lpk/v1
https://www.protocols.io/view/aggregation-propensity-of-synuclein-seeds-in-prima-c7ymzpu6
Arpine Sokratian
TITLE: Aggregation propensity of α-synuclein seeds in primary neurons AUTHORS: Arpine Sokratian [DESCRIPTION] This protocol describes the details of the treatment of primary hippocampal culture with α-synuclein fibrils or monomer protein. It also includes details of the collection of cell lysates for immunoblotting or...
["Prepare primary hippocampal neurons according to protocol", "At DIV21 for 14 days of incubation or at DIV14 for 7 days of incubation culture media should be removed - aspirate the media", "Here are options to use the prepared cultures for the western-blot analysis or ELISA (step-case) or for the immunofluorescence", ...
31,260
Environmental DNA (eDNA) metabarcoding protocol for fish species
1
dx.doi.org/10.17504/protocols.io.bar4id8w
https://www.protocols.io/view/environmental-dna-edna-metabarcoding-protocol-for-bar4id8w
Omneya Ahmed, Tomas Larsson, Mats Töpel, Alexander Eiler
TITLE: Environmental DNA (eDNA) metabarcoding protocol for fish species AUTHORS: Omneya Ahmed, Tomas Larsson, Mats Töpel, Alexander Eiler [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"> Environmental DNA metabarcoding universal primers targeting the hypervariable region of the 12S rRNA gene </div><...
["DNA extraction can be performed using Qiagen DNeasy power water sterivex kit. The quality of the extracted DNA was estimated using Nanodrop. \n\nQiagen DNeasy power water sterivex kit: https://www.qiagen.com/se/resources/resourcedetail?id=c5fe7d5f-070a-4ebe-ac04-4bbf05a13e91&lang=en", "Perform the first PCR (triplica...
45,186
GM Covid-19 saliva test (v2)
4
null
https://www.protocols.io/view/gm-covid-19-saliva-test-v2-bqdams2e
TITLE: GM Covid-19 saliva test (v2) AUTHORS: [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">GM Covid-19 saliva test is a RT-PCR test intended for qualitative detection of nucleic acids from SARS-CoV-2 in saliva specimen. The test is used for screening purpose under class of "research used only" (...
["[Setup reaction]\nPrepare PCR master mixPrepare 100 reactions by adding 1ml Buffer into 1ml Enzyme. Aliquot 20ul into each PCR tube.", "[Setup reaction]\nSaliva-PrepSaliva is self-collected into any sterile tubes with screw cap and wide opening (Sanitize hands before and after saliva collection); Use disposable trans...
21,200
Structured Interview for Protocol Use
null
dx.doi.org/10.17504/protocols.io.yxqfxmw
null
Drew Gitomer
TITLE: Structured Interview for Protocol Use AUTHORS: Drew Gitomer [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is a modification of Palinkas et al SIEU. Based on a lot of feelings and intuitions we thought that a 4 point scale would be super.</div><div class = "text-block">This article des...
[]
93,161
ssDNA2.0: Fill-in mix
3
dx.doi.org/10.17504/protocols.io.dm6gp3111vzp/v1
https://www.protocols.io/view/ssdna2-0-fill-in-mix-c68hzht6
Sarah Nagel, Anna Schmidt, Matthias Meyer
TITLE: ssDNA2.0: Fill-in mix AUTHORS: Sarah Nagel, Anna Schmidt, Matthias Meyer [DESCRIPTION] Protocol for the preparation of Fill-in mix for automated single-stranded DNA library preparation using the ssDNA2.0 method (Gansauge et al. 2020). References Gansauge, M.-T., Aximu-Petri, A., Nagel, S., & Meyer, M. (2020)....
[]
null
null
null
dx.doi.org/10.17504/protocols.io.invcde6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>We propose a Single-Strand Conformation Polymorphism (SSCP) methodology to recognize each species as an effective tool in <em>Diatraea</em> species identification.</p> [STEPS] ?. ?. ?.
[]
66,472
Measurement of dissolved carbohydrate
6
dx.doi.org/10.17504/protocols.io.bp2l6168zvqe/v1
https://www.protocols.io/view/measurement-of-dissolved-carbohydrate-cc6gszbw
Ying-Yu Hu, Zoe V. Finkel
TITLE: Measurement of dissolved carbohydrate AUTHORS: Ying-Yu Hu, Zoe V. Finkel [DESCRIPTION] Here we describe a protocol to measure the dissolved carbohydrate, including total dissolved monosaccharides and total dissolved polysaccharides. For total dissolved carbohydrate measurement, freeze-dried dissolved carbohydr...
["[Sample collection] GFF filter is combusted for 240 min at 450 °C \nGlass filter holder is combusted for 120 min at 500 °C \nGlass filter funnel, flask and 10 mL centrifuge tubes are combusted for 360 min at 500 °C \n \nTube caps are acid-washed.", "[Sample collection] Filter microalgae sample and collect the fi...
49,543
Procedure for measuring extraction efficiency
1
null
https://www.protocols.io/view/procedure-for-measuring-extraction-efficiency-bumfnu3n
Krista Longnecker, Gretchen Swarr
TITLE: Procedure for measuring extraction efficiency AUTHORS: Krista Longnecker, Gretchen Swarr [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Capturing dissolved organic matter from seawater using solid phase extraction is not 100% efficient. We use this protocol to quantify the amount of organic ...
["[Procedure for measuring extraction efficiency]\nThis protocol starts with the eluent from a solid phase extraction resin. Dry down eluents in vials (all the way down, no solvent remaining!).\n8 mL", "[Procedure for measuring extraction efficiency]\nWhile drying down prepare EPA vials with MQ water and HCl.\n40 mL...
40,086
Database Preparation and Search for Protein Identification
3
dx.doi.org/10.17504/protocols.io.bjdwki7e
https://www.protocols.io/view/database-preparation-and-search-for-protein-identi-bjdwki7e
avinash.kale
TITLE: Database Preparation and Search for Protein Identification AUTHORS: avinash.kale [STEPS]
[]
83,246
Chloroform-free DNA Extraction - Ammonium Acetate Precipitation Method
4
dx.doi.org/10.17504/protocols.io.x54v9pekzg3e/v1
https://www.protocols.io/view/chloroform-free-dna-extraction-ammonium-acetate-pr-cvinw4de
NERC Environmental Omics Facility (NEOF) Visitor Facility
TITLE: Chloroform-free DNA Extraction - Ammonium Acetate Precipitation Method AUTHORS: NERC Environmental Omics Facility (NEOF) Visitor Facility [DESCRIPTION] A chloroform-free, cost-effective DNA extraction method for a variety of sample types. [STEPS] 1. Add 250 µL Digsol buffer (see Materials for recipe) and 10 ...
["Add 250 µL Digsol buffer (see Materials for recipe) and 10 µL to a lablelled 1.5ml tube.", "Tissue: Cut into small pieces (<1cm2) with a sterile razor blade on a sterile glass plate before adding to the 1.5mL tube.\nBlood: Centrifuge blood sample at 13,000rpm for about 1 min (to pellet sample).\nRemove sample from ...
75,201
Implant Surgery: Chronic recoverable Neuropixels in mice
1
null
https://www.protocols.io/view/implant-surgery-chronic-recoverable-neuropixels-in-cmn9u5h6
Emily A Aery Jones
TITLE: Implant Surgery: Chronic recoverable Neuropixels in mice AUTHORS: Emily A Aery Jones [DESCRIPTION] This protocol collection explains how to build a low-cost, lightweight system to implant Neuropixels 1.0 probes into mice, record during freely moving behavior, then recover the probe for future use. This protocol...
["[Prepare surgical tools and field] Sterilize tips of metal instruments, ground screw, and headbar in autoclave 25 min or hot bead sterilizer 5 s . Disinfect cotton swabs, toothpick, and a weigh boat under UV light 30 min .", "[Prepare surgical tools and field] Disinfect surgical field with 10% bleach. Lay absorbent ...
62,780
COI library preparation for Illumina MiSeq eDNA metabarcoding - rocky intertidal seawater samples
1
dx.doi.org/10.17504/protocols.io.kxygxzordv8j/v1
https://www.protocols.io/view/coi-library-preparation-for-illumina-miseq-edna-me-b9i4r4gw
Mary McElroy
TITLE: COI library preparation for Illumina MiSeq eDNA metabarcoding - rocky intertidal seawater samples AUTHORS: Mary McElroy [DESCRIPTION] This protocol describes library preparation for COI amplicon metabarcoding with the Illumina MiSeq system. Seawater samples were collected from California rocky intertidal habita...
["[Amplicon PCR] Dilute primers to 5 uM in PCR water. Dilute BSA to 4 mg/ml in PCR water. Set up triplicate PCRs for each sample.", "[Amplicon PCR] Perform PCRs in 20 ul volumes with the following reaction chemistry:\n\n10 ul 2X AmpliTaq Gold 360 Master Mix (Applied Biosystems)\n0.75 ul 4 mg/ml bovine serum albu...
85,828
Basketball Exercise
1
null
https://www.protocols.io/view/basketball-exercise-cx3cxqiw
Ziyong Ma
TITLE: Basketball Exercise AUTHORS: Ziyong Ma [DESCRIPTION] A simple basketball training plan. [GUIDELINES] Just try your best to finish it [STEPS] SECTION: Dribble Exercise 2. Ball Rubs 1 x 10 s SECTION: Dribble Exercise 3. Single Leg Wraps -Left 1 x 20 s -Right 1 x 20 s SECTION: Dribble Exercise 4. 3-3-3 Drib...
["[Dribble Exercise] Ball Rubs\n1 x 10 s", "[Dribble Exercise] Single Leg Wraps\n-Left 1 x 20 s \n-Right 1 x 20 s", "[Dribble Exercise] 3-3-3 Dribble Drill\n 2 x 20 s \nPS: with variations", "[Dribble Exercise] Cone Dribbling\n1 x 3 min \n-with variations", "[Shooting Exercises] Form Shooting (one hand)\n-Left 1 x 20 s...
87,737
TST Nuclei Isolation with GentleMACS - 220301
4
dx.doi.org/10.17504/protocols.io.dm6gp39z1vzp/v1
https://www.protocols.io/view/tst-nuclei-isolation-with-gentlemacs-220301-czwzx7f6
Sébastien Vigneau
TITLE: TST Nuclei Isolation with GentleMACS - 220301 AUTHORS: Sébastien Vigneau [DESCRIPTION] This protocol describes the process of nuclei isolation from frozen tissue. The protocol has been applied to frozen melanoma, breast, and lung metastases for the Human Tumor Atlas Network (HTAN) single-nuclei RNA-seq preparat...
["Buffer Preparation\n\nPrepare the necessary buffers and solutions as outlined below:\n\n2x ST (50 mL stock solution can be prepared ahead of time and stored at room temperature)\n \n\nTST (2 mL should be prepared for each tissue sample)\n*10% Tween-20 can be prepared ahead of time and stored at 4ºC\n \n\nReagent\n\...
50,930
LPCA
3
dx.doi.org/10.17504/protocols.io.bvysn7we
https://www.protocols.io/view/lpca-bvysn7we
Kizito Ndihokubwayo, Jean Uwamahoro, Irénée Ndayambaje, Michael Ralph
TITLE: LPCA AUTHORS: Kizito Ndihokubwayo, Jean Uwamahoro, Irénée Ndayambaje, Michael Ralph [DESCRIPTION] <div class = "text-blocks"></div> [STEPS]
[]
88,717
Barcode Composition by Overlap-Extension PCR
1
dx.doi.org/10.17504/protocols.io.ewov1q2p7gr2/v2
https://www.protocols.io/view/barcode-composition-by-overlap-extension-pcr-c2vmye46
Mathew Chu
TITLE: Barcode Composition by Overlap-Extension PCR AUTHORS: Mathew Chu [DESCRIPTION] Traditionally, DNA barcodes are synthesised as random oligonucleotides. However, this leads to uncertainty regarding the ground truth of barcode sequences in the experimental setting. Without reference sequences, it is impossible to ...
["[Single Stranded DNA Pools for Combinatorial Assembly] ssDNA oligos for combinatorial assembly can be ordered as a pool (oPool). For a final barcode of n units, each with m diversity, order a set of m different barcode sequences for each unit:\n \nwhere unit i (1 ≤ i ≤ n) consists of m barcodes flanked by left (L) an...
19,558
DOI document
null
dx.doi.org/10.17504/protocols.io.xcefite
null
Andrew Khramchenkov
TITLE: DOI document AUTHORS: Andrew Khramchenkov [STEPS] ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.reid3ce
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>On June 14-17, 2018, the 23rd EHA conference was held in Sweden-Stockholm. The heavy research in the blood field took place. The development of new drugs in the field of acute leukemia has progressed rapidly. Among them, antibody-drug conjugates(<a href="https://www.creative-...
[]
98,223
Fiber Photometry Protocol
0
dx.doi.org/10.17504/protocols.io.eq2lyw39qvx9/v1
https://www.protocols.io/view/fiber-photometry-protocol-db6p2rdn
Sasha Burwell
TITLE: Fiber Photometry Protocol AUTHORS: Sasha Burwell [DESCRIPTION] This protocol details collection of fiber photometry from VTA dopamine neurons. [GUIDELINES] Recordings collected with a Tucker-Davis Technologies RZ10X and TDT Synapse software. [STEPS] SECTION: On every day of behavior (see Behavior protocol): 1...
["[On every day of behavior (see Behavior protocol):] Photobleach the mono fiberoptic patchcords (Doric Lenses Inc., MFP_400/430/1100-0.57_1mm_FCM-MF1.25_LAF) (Synapse -> Preview Mode -> Fiber Bleaching) for at least 60 min, and up to 240 min, in both the 415nm and 465nm channels.\n\nThis can be done the night before t...
53,104
Illumina DNA Prep (M) Tagmentation Library Preparation for use on an Illumina MiSeq Sequencer
1
dx.doi.org/10.17504/protocols.io.bx4qpqvw
https://www.protocols.io/view/illumina-dna-prep-m-tagmentation-library-preparati-bx4qpqvw
Julie Haendiges, Narjol Gonzalez-Escalona, Ruth Timme, Maria Balkey
TITLE: Illumina DNA Prep (M) Tagmentation Library Preparation for use on an Illumina MiSeq Sequencer AUTHORS: Julie Haendiges, Narjol Gonzalez-Escalona, Ruth Timme, Maria Balkey [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This procedure outlines the protocol for whole genome sequencing of bacter...
["[Dilute and Tagment Input DNA]\nBring BLT (stored in refrigerator) and TB1 (stored in freezer) to room temperature. Ensure that BLT is stored upright at all times, so that the beads remain submerged in thebuffer", "[Dilute and Tagment Input DNA]\nLabel a 96-well PCR plate with the Run ID.", "[Dilute and Tagment Inpu...
26,574
RNA isolation from suspended animal cells, cDNA library construction, and RNA-Seq for gene expression analysis
1
dx.doi.org/10.17504/protocols.io.57ng9me
https://www.protocols.io/view/rna-isolation-from-suspended-animal-cells-cdna-lib-57ng9me
Tomoko Matsuda
TITLE: RNA isolation from suspended animal cells, cDNA library construction, and RNA-Seq for gene expression analysis AUTHORS: Tomoko Matsuda [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is for RNA Isolation from suspended Chinese hamster ovary (CHO) cells. Wash cells once in 1× PBS...
["Cells Maintenance", "CHO-S cells were grown in 20 ml of Gibco CD-CHO medium (Thermo Fisher Scientific, Waltham, MA, USA) containing 400 µL of 200 mM L-Alanyl-L-glutamine (final concentration: 4 mM; Sigma-Aldrich, St. Louis, MI, USA) and 40 µL of Gibco Anti-Clumping Agent (final concentration, 0.2% (v/v); Thermo Fishe...
74,126
Sugar Beet Tissue Collection for Genome Assembly and Annotation with Long Read Sequencing
4
null
https://www.protocols.io/view/sugar-beet-tissue-collection-for-genome-assembly-a-ckmnuu5e
olivia.todd
TITLE: Sugar Beet Tissue Collection for Genome Assembly and Annotation with Long Read Sequencing AUTHORS: olivia.todd [DESCRIPTION] This protocol outlines the various types of tissue collection needed to follow the USDA-ARS Dorn lab protocol for Beta vulgaris crop wild relative genome sequencing. The annotation pipeli...
["Follow the protocol below as written to isolate plant nuclei from 1-5g of dark treated tissue.\nhttps://15a13b02-7dac-4315-baa5-b3ced1ea969d.filesusr.com/ugd/5518db_90f751986a4a4e7bbc8e648d467507d2.pdf?index=true", "Check for quality and presence of necessary DNA size for sequencing (>15kb) using a Pippin Pulse, runn...
null
null
null
dx.doi.org/10.17504/protocols.io.iiwccfe
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>The procedures to determine the half-lives of proteins by pulse-chase analysis and cycloheximide blocking are well established. In the case of oligomeric proteins, these methods do not differentiate whether the stabilities of the monomeric versus oligomeric forms of the prote...
[]
38,787
Plasmid Extraction
4
null
https://www.protocols.io/view/plasmid-extraction-bh5bj82n
Hung Liang Pai, Huan Jui Chang
TITLE: Plasmid Extraction AUTHORS: Hung Liang Pai, Huan Jui Chang [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is used for extracting plasmid from E. coli DH5-Alpha strains. </div></div> [STEPS] ?. [Preparation] Set the dry bath incubator at 37 °C ?. [Preparation] Prepare Presto™ M...
["[Preparation]\nSet the dry bath incubator at\n37 °C", "[Preparation]\nPrepare Presto™ Mini Plasmid Kit", "[Preparation]\nTake out the centrifuge tube containing medium with bacteria that has been cultured overnight.", "[Protocol]\nCentrifuge the centrifuge tube under this condition - .\nCentrifuge: 15000 34, 6 min"...
19,777
U Cinn - Energy Expenditure Measurements
null
dx.doi.org/10.17504/protocols.io.xi9fkh6
null
Patrick Tso
TITLE: U Cinn - Energy Expenditure Measurements AUTHORS: Patrick Tso [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span></div><div class = "text-block"> Oxygen consumption and carbon dioxide production is measured using indirect calorimetry. The Columbus...
["Energy Expenditure by Oxymax Protocol (Columbus Instruments)1. Take Body Composition via NMR before you put animals in this experiment.\n2. Set up cages (this can be done at any time before starting an experiment)\n a. Weigh animals and place into Oxymax chambers\n b. Supply food and water\n c. Make sure c...
null
null
null
dx.doi.org/10.17504/protocols.io.fctbiwn
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?.
[]
71,507
Monkeypox virus multiplexed PCR amplicon sequencing (PrimalSeq) 
1
dx.doi.org/10.17504/protocols.io.5qpvob1nbl4o/v3
https://www.protocols.io/view/monkeypox-virus-multiplexed-pcr-amplicon-sequencin-ch3tt8nn
Nicholas F.G. Chen*, Luc Gagne*, Matthew Doucette, Sandra Smole, Erika Buzby, Joshua Hall, Stephanie Ash, Rachel Harrington, Seana Cofsky, Selina Clancy, Curtis J Kapsak, Joel Sevinsky, Kevin Libuit, mallery.breban, Chrispin Chaguza, Nathan D. Grubaugh, Daniel J. Park, Glen R. Gallagher#, Chantal B.F. Vogels#
TITLE: Monkeypox virus multiplexed PCR amplicon sequencing (PrimalSeq)  AUTHORS: Nicholas F.G. Chen*, Luc Gagne*, Matthew Doucette, Sandra Smole, Erika Buzby, Joshua Hall, Stephanie Ash, Rachel Harrington, Seana Cofsky, Selina Clancy, Curtis J Kapsak, Joel Sevinsky, Kevin Libuit, mallery.breban, Chrispin Chaguza, N...
["[Dilute and Pool Primers] Reagents:", "[Dilute and Pool Primers] If not already done, separate odd and even numbered primer pairs into two separate boxes. These will constitute the two pools", "[Dilute and Pool Primers] Label 164, 8-strip tubes with the corresponding odd-numbered primer name (e.g. 3 left)", "[Dilute ...
43,141
Protocol 2: PCR Wet Lab
5
null
https://www.protocols.io/view/protocol-2-pcr-wet-lab-bnddma26
TITLE: Protocol 2: PCR Wet Lab AUTHORS: [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. DNA ExtractionDNA Extraction: Saliva (Bento lab) This protocol extracts DNA from Saliva using a microcentrifuge and thermocycler. The microcentrifuge separates the cells in the sample into a pellet at the bottom of ...
["DNA ExtractionDNA Extraction: Saliva (Bento lab) This protocol extracts DNA from Saliva using a microcentrifuge and thermocycler. The microcentrifuge separates the cells in the sample into a pellet at the bottom of the microcentrifuge tube. Extra liquid is drawn out of the microcentrifuge tube. Cells are then heated...
79,067
Single-cell dissociation of Drosophila melanogaster pupal tarsi
4
dx.doi.org/10.17504/protocols.io.x54v9dzbmg3e/v1
https://www.protocols.io/view/single-cell-dissociation-of-drosophila-melanogaste-crf3v3qn
Ben R. Hopkins, Olga Barmina, Artyom Kopp
TITLE: Single-cell dissociation of Drosophila melanogaster pupal tarsi AUTHORS: Ben R. Hopkins, Olga Barmina, Artyom Kopp [DESCRIPTION] This protocol outlines a step-by-step guide to generating single-cell suspensions of Drosophila melanogaster pupal tarsi for use in 10x single-cell transcriptome profiling. This proto...
["[Collecting, sexing, and ageing pupae] Collect white prepupae. Individuals should meet the P1 aging criteria laid out by Bainbridge and Bownes (1981): the pupae should be white or cream coloured, have stopped moving completely, and display everted anterior spiracles.", "Identify individuals of the correct sex. Place ...
49,128
Initial Rapid Pathology Assessment of Kidney Tissue
1
dx.doi.org/10.17504/protocols.io.bt8gnrtw
https://www.protocols.io/view/initial-rapid-pathology-assessment-of-kidney-tissu-bt8gnrtw
Jamie Allen, Carrie Romer, Elizabeth Neumann, Maya Brewer, Haichun Yang, Jeff Spraggins, Danielle Gutierrez
TITLE: Initial Rapid Pathology Assessment of Kidney Tissue AUTHORS: Jamie Allen, Carrie Romer, Elizabeth Neumann, Maya Brewer, Haichun Yang, Jeff Spraggins, Danielle Gutierrez [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Scope:</div><div class = "text-block">Prepare formalin fixed tissue with fr...
["Tissue is embedded in protocol: dx.doi.org/10.17504/protocols.io.br4fm8tn", "Section samples at 5 µm on a microtome.", "PAS stain tissue sections with protocol: dx.doi.org/10.17504/protocols.io.buaknscw", "Scan slides with brightfield scanner (Leica) and save as .tiff or .jpg", "Place saved images on QuPath for analy...
32,472
PCR amplification of long GC-rich DNA targets
1
dx.doi.org/10.17504/protocols.io.bbxyippw
https://www.protocols.io/view/pcr-amplification-of-long-gc-rich-dna-targets-bbxyippw
Nadia Assal, Min Lin
TITLE: PCR amplification of long GC-rich DNA targets AUTHORS: Nadia Assal, Min Lin [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol allows the amplification of long and high GC content genes by PCR without the need for many optimizations. In this protocol, the</span><span style = ...
["Extract M. bovis genomic DNA using according to the manufacturer’s instructions for Gram-positive bacteria. Use 5 ng/µl of genomic DNA was used in each reaction", "Prepare the master mix for the polymerase enzyme as follows: AB1Componentvolume in µl2Milliq Water29.53Buffer104DNTPs&nbsp; (2.5mM)45DMSO2.56Primers Mix\...
25,251
Marchantia spores sterilisation
null
dx.doi.org/10.17504/protocols.io.4wbgxan
null
Linda Silvestri, Eftychis Frangedakis, Marius Rebmann, Susana Sauret-Gueto
TITLE: Marchantia spores sterilisation AUTHORS: Linda Silvestri, Eftychis Frangedakis, Marius Rebmann, Susana Sauret-Gueto [STEPS] ?. Add one Milton tablet into 25 mL of sterile water to prepare the "sterilization solution". ?. Add the sporangia into a 1.5 mL centrifuge tube and add 0.5 mL of sterilization solution in...
["Add one Milton tablet into 25 mL of sterile water to prepare the \"sterilization solution\".", "Add the sporangia into a 1.5 mL centrifuge tube and add 0.5 mL of sterilization solution into the tube (B in Figure)", "Use sterile metal tweezers to crush the sporangia (C in Figure).", "Place a 40 μM cell strainer into a...
31,227
Optical mapping preps for Petunia spp.
null
dx.doi.org/10.17504/protocols.io.baq3idyn
null
Elena Hilario
TITLE: Optical mapping preps for Petunia spp. AUTHORS: Elena Hilario [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Optical mapping technologies assist the assembly of large and complex genomes. Although protocols for preparing this kind of samples are now conveniently available as kits, some plant...
["[Tissue Fixation]\nRemove ~ and place them in a 50 mL Falcon tube containing ~. Attach the green screened cap and slowly add more Fixing solution (total volume 60 mL).Remove all the air bubbles by tapping the tube but keeping it upright. Close the tube with its own cap and incubate on ice.\nDo the tissue fixation a...
63,409
Lumina Luxe Face Cream Reviews Best Anti Aging Cream for Glamorous Look!
3
dx.doi.org/10.17504/protocols.io.81wgb69p1lpk/v1
https://www.protocols.io/view/lumina-luxe-face-cream-reviews-best-anti-aging-cre-b96rr9d6
Lumina Luxe Face Cream
TITLE: Lumina Luxe Face Cream Reviews Best Anti Aging Cream for Glamorous Look! AUTHORS: Lumina Luxe Face Cream [DESCRIPTION] Product Name: Lumina Luxe Face Cream Category: FACE SKIN CARE CREAM Price: $8 (for 1 box) Official Website: Click Here [STEPS]
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null
null
null
dx.doi.org/10.17504/protocols.io.cqqvvv
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?.
[]
20,958
Cell Hashing
null
dx.doi.org/10.17504/protocols.io.yp6fvre
null
Brenton Paolella
TITLE: Cell Hashing AUTHORS: Brenton Paolella [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is for performing Cell Hashing only. </div><div class = "text-block">Sample multiplexing and super-loading on single cell RNA-sequencing platforms.</div><div class = "text-block"><a style = "t...
["[Cell staining for Drop-seq or 10x Genomics]\nObtain all single cell suspensions from different samples/conditions that will be multiplexed in the run. Keep samples in separate tubes until after cell hashing and shortly before loading cells into the single cell RNA-seq instrument. When aiming to super-load the same s...
49,233
Human intestinal cell dissociation suitable for multi-omics single-cell assays
4
dx.doi.org/10.17504/protocols.io.bubrnsm6
https://www.protocols.io/view/human-intestinal-cell-dissociation-suitable-for-mu-bubrnsm6
Astrid Kosters, Junkai Yang, Ann Dodd, Mackenzie White, Greg Gibson, Subra Kugathasan, Peng Qiu, Eliver Ghosn
TITLE: Human intestinal cell dissociation suitable for multi-omics single-cell assays AUTHORS: Astrid Kosters, Junkai Yang, Ann Dodd, Mackenzie White, Greg Gibson, Subra Kugathasan, Peng Qiu, Eliver Ghosn [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">We developed a protocol for the preparation ...
["Chop tissue into small pieces with a razor blade in def-RPMI/3%NBCS/EDTA/DTT media in a 6 cm dish, on ice.", "Transfer tissue solution to 15 mL conical (use 1 mL pipet).", "Vortex the tissue solution, and incubate at RT for 5 min, vortexing intermittently.", "Fill the 15 mL conical with def-RPMI/3%NBCS.", "Spin for 5...
91,233
Cartilage staining
4
null
https://www.protocols.io/view/cartilage-staining-c5b9y2r6
Satheeswaran Balasubramanian, Ekambaram Perumal
TITLE: Cartilage staining AUTHORS: Satheeswaran Balasubramanian, Ekambaram Perumal [DESCRIPTION] The Alcian Blue staining technique is widely used among developmental biologists to observe the embryonic development of cartilage and bone structures in embryos and complete zebrafish larvae. Alcian blue is a positively c...
["[Larval fixation] At the desired stage, the larvae for staining are taken and washed once in PBS for 5 minutes.", "[Larval fixation] The larvae were euthanized using the cold shock method (Keep at 4oC for 5-10 minutes).", "[Larval fixation] Transfer the euthanized larvae into 4% PFA and keep it in the rocker for 2 ho...
55,026
NAN KB Demo: Data download
5
null
https://www.protocols.io/view/nan-kb-demo-data-download-bzysp7we
Abby Moore
TITLE: NAN KB Demo: Data download AUTHORS: Abby Moore [DESCRIPTION] The purpose of this protocol is to describe how data may be downloaded from "our project." The scope of this protocol extends to any instance where a user must download data from a spectrometer that is a part of "our project." [BEFORE_START] For t...
["[Data download] Select the appropriate case below to proceed.", "[Data download] Open a terminal in", "[Data download] Copy your data from the spectrometer using the following command:", "[Data download] You will be asked for your account password - type this in and hit enter, your download should commence immediatel...
58,022
Differentiation of iPSC into Microglia-Like Cells (iMGL)
1
dx.doi.org/10.17504/protocols.io.q26g7bwqklwz/v3
https://www.protocols.io/view/differentiation-of-ipsc-into-microglia-like-cells-b4weqxbe
Abhirami Kannan Iyer, Emma Danhash, Fabia Filipello, Jacob Marsh, Rj Martinez, Celeste M M. Karch
TITLE: Differentiation of iPSC into Microglia-Like Cells (iMGL) AUTHORS: Abhirami Kannan Iyer, Emma Danhash, Fabia Filipello, Jacob Marsh, Rj Martinez, Celeste M M. Karch [DESCRIPTION] This protocol outlines the derivation of Hematopoietic Progenitor Cells and differentiation of iMGLs using iPSC cultures. This pr...
["[iPSCs Culture] Thaw and culture iPSC line per the following protocol:", "[iPSCs Aggregate Plating] Once iPSCs are 70-80% confluent in 2-3 wells of a 6-well tissue culture plate, passage and plate the iPSCs as aggregates", "[iPSCs Aggregate Plating] Coat a 6-well tissue culture plate with Matrigel for a least 60 min ...
92,335
Patient PBMC flow cytometry
4
dx.doi.org/10.17504/protocols.io.n92ldm688l5b/v1
https://www.protocols.io/view/patient-pbmc-flow-cytometry-c6epzbdn
rebeccawallings
TITLE: Patient PBMC flow cytometry AUTHORS: rebeccawallings [DESCRIPTION] Patient PBMC flow cytometry [STEPS] 1. 1 x 106 PBMCs were taken for flow cytometry and transferred to a v-bottom 96-well plate (Sigma, CLS3896-48EA) and centrifuged at 300x g for 5 minutes at 4°C. Cells were resuspended in 50 µL of PBS contain...
["1 x 106 PBMCs were taken for flow cytometry and transferred to a v-bottom 96-well plate\n(Sigma, CLS3896-48EA) and centrifuged at 300x g for 5 minutes at 4°C.\n\nCells were resuspended in 50 µL of PBS containing diluted fluorophore-conjugated antibodies and incubated in the dark at 4°C for 20 minutes. Cells were cent...
42,463
ELISA for assessing the burden of Neutrophil Extracellular Traps (NETs) in clinical serum samples.
4
dx.doi.org/10.17504/protocols.io.bmp7k5rn
https://www.protocols.io/view/elisa-for-assessing-the-burden-of-neutrophil-extra-bmp7k5rn
kathryn.hally
TITLE: ELISA for assessing the burden of Neutrophil Extracellular Traps (NETs) in clinical serum samples. AUTHORS: kathryn.hally [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol steps through the ELISA procedure for detecting three NET-specific biomarkers: 1) myeloperoxidase-DNA (MPO-D...
["[Solutions to be prepared for ELISAs.]\nFor Day 1:Coating buffer:\n[1x PBS]", "[ELISA day 2.]\nThoroughly remove the capture antibody by flicking off the solution and banging each plate face down on paper towels. Add to each well and incubate for . Flick and bang again. Repeat another four times (five washes in to...
78,863
Pathogen-Oriented Low-cost Assembly &amp; Re-sequencing on Opentrons (POLARtron): An automation-friendly highly sensitive and high-throughput SARS-CoV-2 diagnostic based on whole genome sequencing
4
dx.doi.org/10.17504/protocols.io.81wgbp8jqvpk/v2
https://www.protocols.io/view/pathogen-oriented-low-cost-assembly-amp-re-sequenc-cq9pvz5n
Per A. Adastra, Neva C. Durand, Namita Mitra, Saul Godinez, Ragini Mahajan, Alyssa Blackburn, Zane Colaric, Joshua W. M. Theisen, David Weisz, Olga Dudchenko, Andreas Gnirke, Suhas S.P. Rao, Parwinder Kaur, Erez Lieberman Aiden, Aviva Presser Aiden
TITLE: Pathogen-Oriented Low-cost Assembly &amp; Re-sequencing on Opentrons (POLARtron): An automation-friendly highly sensitive and high-throughput SARS-CoV-2 diagnostic based on whole genome sequencing AUTHORS: Per A. Adastra, Neva C. Durand, Namita Mitra, Saul Godinez, Ragini Mahajan, Alyssa Blackburn, Zane Colaric...
["[RNA Extraction] For each saliva sample recieved add equal volume saliva and 2X DNA/RNA Shield from the Quick-DNA/RNA Viral magbead kit and vortex. Centrifuge the samples at 500 rpm, 5 min to bring down debris.", "[RNA Extraction] Without disturbing the pellet, transfer 25 µL of saliva sample and 1X DNA/RNA to the ...
29,287
High-quality RNA purification with on-column DNase treatment from tissue specimens
null
dx.doi.org/10.17504/protocols.io.8ufhwtn
null
Magda Bletsa, Antonios Fikatas, Yiqiao Li, Sophie Gryseels, Jan Felix Drexler, Philippe Lemey
TITLE: High-quality RNA purification with on-column DNase treatment from tissue specimens AUTHORS: Magda Bletsa, Antonios Fikatas, Yiqiao Li, Sophie Gryseels, Jan Felix Drexler, Philippe Lemey [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol can be used for total RNA purification from ...
["[Sample homogenisation]\nAdd RLT Buffer to the Precellys lysate tubes.\n600 µl", "[Sample homogenisation]\nExcise a lentil-sized piece of tissue (maximum amount of for RNAlater stabilized tissues and for fresh or frozen tissues) and transfer it quickly to the lysate tubes. Make sure that all tissues are immersed in...
39,590
Direct wastewater RNA capture and purification via the "Sewage, Salt, Silica and SARS-CoV-2 (4S)" method
4
dx.doi.org/10.17504/protocols.io.biwekfbe
https://www.protocols.io/view/direct-wastewater-rna-capture-and-purification-via-biwekfbe
Oscar Whitney, Basem Al-Shayeb, Alex Crits-Cristoph, Mira Chaplin, Vinson Fan, Hannah Greenwald, Adrian Hinkle, Rose Kantor, Lauren Kennedy, Anna Maurer, Robert Tjian, Kara L. Nelson, UC Berkeley Wastewater-based epidemiology consortium
TITLE: Direct wastewater RNA capture and purification via the "Sewage, Salt, Silica and SARS-CoV-2 (4S)" method AUTHORS: Oscar Whitney, Basem Al-Shayeb, Alex Crits-Cristoph, Mira Chaplin, Vinson Fan, Hannah Greenwald, Adrian Hinkle, Rose Kantor, Lauren Kennedy, Anna Maurer, Robert Tjian, Kara L. Nelson, UC Berkeley Was...
["[Sample preparation, RNA preservation and particle lysis]\nSpike a known volume and titer of bovine coronavirus (bCoV) into the wastewater sample as a recovery efficiency control. Agitate sample to fully mix bCoV or other spiked-in controls with the wastewater sample.\nOther recovery controls can be used instead of b...
81,551
High molecular weight DNA extraction for marine macroalgal tissue
4
dx.doi.org/10.17504/protocols.io.14egn2dnpg5d/v1
https://www.protocols.io/view/high-molecular-weight-dna-extraction-for-marine-ma-ctvpwn5n
Malia Moore, Taylor S. Steele
TITLE: High molecular weight DNA extraction for marine macroalgal tissue AUTHORS: Malia Moore, Taylor S. Steele [DESCRIPTION] This protocol details high molecular weight DNA extraction for marine macroalgal tissue. Marine macroalgae contain a variety of unique cell wall components including sulfated polysaccharides an...
["[Lyophilizing algal tissue] Flash-freeze algal tissue in liquid nitrogen (target ≥5 g wet tissue).", "[Setting up the DNA extraction] Prepare desired volume of Carlson lysis buffer (100 millimolar (mM) Tris-HCl, pH 9.5, 2% CTAB, 1.4 Molarity (M) NaCl, 1% PEG 8000, 20 millimolar (mM) EDTA) and mix 1440 min on a magnet...
29,666
Enrichment of a specific polyadenylated RNA for nanopore direct RNA sequencing (RNA SPACE)
null
dx.doi.org/10.17504/protocols.io.88ahzse
null
Paul MK. Gordon
TITLE: Enrichment of a specific polyadenylated RNA for nanopore direct RNA sequencing (RNA SPACE) AUTHORS: Paul MK. Gordon [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This RNA Sequence Picking After Cutting Enzymatically (RNA SPACE) protocol is intended to enrich for a specific polyadenyla...
["[Oligonucleotide design]\nHave the three oligos synthesized (RE oligo, RTA Oligos A & B), by designing your own using the open source software and your reference data.", "[Enzymatic cleavage of target RNA]\nIn the same tube, perform 3h or overnight restriction digest according to the NEB protocol for your enzyme, e.g...
83,141
SynBot Protocols
2
dx.doi.org/10.17504/protocols.io.3byl4qewjvo5/v1
https://www.protocols.click/view/synbot-protocols-cvfdw3i6
Justin T Savage, Juan Ramirez, Dolores Irala, Cagla Eroglu
TITLE: SynBot Protocols AUTHORS: Justin T Savage, Juan Ramirez, Dolores Irala, Cagla Eroglu [DESCRIPTION] Protocols related to the paper SynBot: An open-source image analysis software for automated quantification of synapses. Includes details of rat neuron and astrocyte culture, in vitro synapse staining, in vivo syna...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.qxfdxjn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>The purinosome has been observed in a broad spectrum of cells, but some studies claim that it is an artefact of the constructs used for visualization or stress granules resulting from the exposure of cells to nutrient-reduced growth media. Both may be true depending on the me...
[]
94,482
MBP-Clu-tail purification from Escherichia coli cells
1
dx.doi.org/10.17504/protocols.io.6qpvr35xzvmk/v1
https://www.protocols.io/view/mbp-clu-tail-purification-from-escherichia-coli-ce-c8hszt6e
Andreas Bracher, F Ulrich Hartl
TITLE: MBP-Clu-tail purification from Escherichia coli cells AUTHORS: Andreas Bracher, F Ulrich Hartl [DESCRIPTION] This protocol details how to efficiently purify the fusion protein Maltose binding protein (MBP)-Clu-tail (204-238) from Escherichia coli. [STEPS] SECTION: His6-Ubiquitin-GFP-Clu-tail expression and cel...
["[His6-Ubiquitin-GFP-Clu-tail expression and cell lysis] Express MBP-Clu-tail in E. coli Bl21 (DE3) codon+RIL cells cultured in 1 L LB Medium containing 2 1655 glucose at 37 °C with 1 millimolar (mM) IPTG during 120 min.", "[His6-Ubiquitin-GFP-Clu-tail expression and cell lysis] Centrifuge culture and keep pellet.", "...
36,796
Coating Slides with Gelatin
null
dx.doi.org/10.17504/protocols.io.bf64jrgw
https://www.protocols.io/view/coating-slides-with-gelatin-bf64jrgw
Allen Institute for Brain Science
TITLE: Coating Slides with Gelatin AUTHORS: Allen Institute for Brain Science [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The protocol describes how 1x3 glass microscope slides are coated with gelatin to improve adherence of fixed adult mouse brain sections or other tissue to slides.</div><div c...
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null
null
null
dx.doi.org/10.17504/protocols.io.c8bzsm
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This is a very simple protocol&nbsp;showing how to extract haemolymph from adult Drosophila melanogaster. (Based on protocols from Sigma Aldrich). [GUIDELINES] <span class="s1"><strong>Materials:</strong></span> <br /><span class="s1">-0.5 &mu;l capillaries with adequate suctio...
[]
47,103
Electroporation of Cas9 protein into human pluripotent stem cells
1
dx.doi.org/10.17504/protocols.io.br87m9zn
https://www.protocols.io/view/electroporation-of-cas9-protein-into-human-pluripo-br87m9zn
Jiuchun Zhang, Harper JW
TITLE: Electroporation of Cas9 protein into human pluripotent stem cells AUTHORS: Jiuchun Zhang, Harper JW [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol describes the electroporation of Cas9 protein into human pluripotent stem cells.</div></div> [STEPS] ?. Add to a sterile 1.5 ml t...
["Add to a sterile 1.5 ml tube. Add . Then add . Pipet up and down to mix. Let it sit at for . This is enough for 2 transfections (== one 6 well).\n[buffer R ]\n[purified Cas9 protein (2mg/ml)]\n[sgRNA]\n0 Room temperature", "While waiting for the Cas9 to bind to sgRNA, individualize cells with Accutase. Neutralize A...
49,968
Human breast tissue dissociation and FACS vs Flowmi processing for scRNA-Seq
4
dx.doi.org/10.17504/protocols.io.bu2qnydw
https://www.protocols.io/view/human-breast-tissue-dissociation-and-facs-vs-flowm-bu2qnydw
Maren Pein, smallya , Quy Nguyen, Jacob Insua-Rodríguez
TITLE: Human breast tissue dissociation and FACS vs Flowmi processing for scRNA-Seq AUTHORS: Maren Pein, smallya , Quy Nguyen, Jacob Insua-Rodríguez [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Single cell RNA-Seq of human breast tissue requires tissue dissociation into viable single cell suspens...
["Initial Tissue PreparationTransfer breast tissue specimen to 150 x 22 mm cell culture dish. Remove medium and wash 3 times with 50-100 mL ice-cold PBS.Optional: If histological analysis is desired, separate tissue pieces and process as desired.", "Mechanical DigestionUsing forceps and scalpels, remove soft, white/yel...