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86,417
Behavior Tracking with Running Wheels
4
null
https://www.protocols.io/view/behavior-tracking-with-running-wheels-cymrxu56
Victoria Vance, Katerina Rademacher, Ken Nakamura
TITLE: Behavior Tracking with Running Wheels AUTHORS: Victoria Vance, Katerina Rademacher, Ken Nakamura [DESCRIPTION] This protocol describes tracking and analyzing behavior in mice using wireless running wheels. Because these running wheels are able to be placed inside an animal's home cage and continuously collect d...
["[Setting Up Wheels] Single-house mice in new cages containing activity wheels at least 3 days prior to beginning drug/vehicle to collect baseline data.", "[Setting Up Wheels] Place clear wheel base holder on cage floor. If the experiment involves providing medicated water, place water bottle in the cage with sufficie...
103,537
Isolation of brain infiltrating lymphocytes
0
dx.doi.org/10.17504/protocols.io.dm6gpzo68lzp/v1
https://www.protocols.io/view/isolation-of-brain-infiltrating-lymphocytes-dhcr32v6
Moustafa Nouh Elemeery, Salix Boulet
TITLE: Isolation of brain infiltrating lymphocytes AUTHORS: Moustafa Nouh Elemeery, Salix Boulet [DESCRIPTION] This protocol details the isolation of brain infiltrating lymphocytes. Splenic lymphocytes are screened to verify the presence/phenotype of CD8 in the periphery. [STEPS] SECTION: Infiltration procedure 1. I...
["[Infiltration procedure] Inject mice intravenously with CD45-FITC 3ug/mice (6 µL from vial/mice) for 3 min-5 min, then euthanized directly by decapitating head to stop blood circulation.", "[Infiltration procedure] Collect the brain for each experimental condition vs control.", "[Infiltration procedure] Dissociate br...
null
null
null
dx.doi.org/10.17504/protocols.io.kr2cv8e
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?.
[]
71,134
Ligation
1
dx.doi.org/10.17504/protocols.io.6qpvr4dmbgmk/v1
https://www.protocols.io/view/ligation-chp6t5re
Georgina Elisa Diego Macías, Ana Belem García González, Irán Alessandra Chaparro Rodríguez, Jair Alexis Gardea Sáenz
TITLE: Ligation AUTHORS: Georgina Elisa Diego Macías, Ana Belem García González, Irán Alessandra Chaparro Rodríguez, Jair Alexis Gardea Sáenz [DESCRIPTION] Protocol for a Ligation with Master Mix Ligase after a restriction digestion protocol had been made. [STEPS] 1. Add5 µL of Master Mix Ligase 2. Add the a quantit...
["Add5 µL of Master Mix Ligase", "Add the a quantity of the DNA vector digested by EcoRI and PstI in a molar ratio 1:3 with the DNA insert", "Add the a quantity of the DNA insert digested by EcoRI and PstI in a molar ratio 3:1 with the DNA vector", "Add water up to 20 µL", "Ligate for 1200 min at 22°C", "Transform with...
38,296
Lentiviral_vector_production_with_PEI
4
null
https://www.protocols.io/view/lentiviral-vector-production-with-pei-bhmyj47w
Kenneth Matreyek, Anna Bruchez
TITLE: Lentiviral_vector_production_with_PEI AUTHORS: Kenneth Matreyek, Anna Bruchez [STEPS] ?. [Preparation] This protocol will require transfecting HEK 293T cells. Make sure you have some 293T cells thawed and growing for at least a couple of days before doing this protocol. Furthermore, prepare stocks of PEI and Di...
["[Preparation]\nThis protocol will require transfecting HEK 293T cells. Make sure you have some 293T cells thawed and growing for at least a couple of days before doing this protocol. Furthermore, prepare stocks of PEI and Diluent if you have not done so already.", "[Transfection]\nPrepare the mixture in a sterile mic...
85,701
TCR sequencing and activation
4
dx.doi.org/10.17504/protocols.io.j8nlkokpxv5r/v1
https://www.protocols.io/view/tcr-sequencing-and-activation-cxxdxpi6
Zorea Jonathan
TITLE: TCR sequencing and activation AUTHORS: Zorea Jonathan [DESCRIPTION] This protocol is a procedure we follow to isolate tumor infiltrating T cells and try to activate them with unique peptides and to characterize their TCR repertoire [STEPS] SECTION: Tumor implantation 1. Grow sufficient amount of cells (5*10^6...
["[Tumor implantation] Grow sufficient amount of cells (5*10^6 per mouse). At the day cells are ready to be inject, thaw a vial of Basement Membrane Extract (Biotest, 3433-010-01) on ice. Keep 200ul tips in -20° as well.", "Once the BME is thawed, trypsinize the cells, count them using trypan blue, and resuspend them i...
38,609
Aspergillus nidulans protoplast isolation for transfections
4
dx.doi.org/10.17504/protocols.io.bhxrj7m6
https://www.protocols.io/view/aspergillus-nidulans-protoplast-isolation-for-tran-bhxrj7m6
Andrew Liu, Jai Denton
TITLE: Aspergillus nidulans protoplast isolation for transfections AUTHORS: Andrew Liu, Jai Denton [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Modified </span><span style = "font-style:italic;">Aspergillus nidulans</span><span> protoplast isolation using Novozyme VinoTaste Pro as an enzyme...
["[Grow Overnight Culutre]\nLate afternoon-evening the day before, grow scraped condispores (two arms from MM complete agar plate, ~ 1 x 108 total spores) in 30ml liquid CM, supplemented with pyridoxine and riboflavin, overnight at 25°C 18-20 hrs or 30°C 11-12 hrs on orbital shaker at 150rpm. Growth can be arrested at ...
27,927
Creation of low-oxygen conditions.
null
dx.doi.org/10.17504/protocols.io.7hxhj7n
null
Shengyi Yan, 宏亮 董
TITLE: Creation of low-oxygen conditions. AUTHORS: Shengyi Yan, 宏亮 董 [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>We use Na</span><span style = "vertical-align:sub;">2</span><span>SO</span><span style = "vertical-align:sub;">3 </span><span>to create low-oxygen conditions in our experiment. ...
["[Changes of dissolved oxygen with time in the LB medium with different concentrations of Na2SO3.]\nPrepare LB medium with different concentrations of Na2SO3.", "[Changes of dissolved oxygen with time in the LB medium with different concentrations of Na2SO3.]\nAB1Na2SO3(100g/L)LB medium20μL20mL320μL20mL440μL20mL5100μL...
19,796
MBARI Environmental DNA (eDNA) extraction using Qiagen DNeasy Blood and Tissue Kit
null
dx.doi.org/10.17504/protocols.io.xjufknw
null
Kristine Walz, Kevan Yamahara, Reiko P Michisaki, Francisco P. Chavez
TITLE: MBARI Environmental DNA (eDNA) extraction using Qiagen DNeasy Blood and Tissue Kit AUTHORS: Kristine Walz, Kevan Yamahara, Reiko P Michisaki, Francisco P. Chavez [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Nucleic acids extraction from the filters using the Qiagen DNeasy Blood and Tissue...
["[Preparation]\nNucleic acids were extracted from the filters using the Qiagen DNeasy Blood and Tissue Kit with some modifications to the manufacturer’s protocol.", "[Preparation]\nPrior to extraction, 0.5 mm and 0.1 mm glass beads (BioSpec Products) were ashed at 500 °C for 5 hours.", "[Preparation]\nBead tubes: 0.25...
91,954
Protocol for fixing and peeling Aedes aegypti embryos
1
dx.doi.org/10.17504/protocols.io.261ged88wv47/v1
https://www.protocols.io/view/protocol-for-fixing-and-peeling-aedes-aegypti-embr-c52sy8ee
William Reid, Geyenna Sterling-Lentsch, Marc S. Halfon
TITLE: Protocol for fixing and peeling Aedes aegypti embryos AUTHORS: William Reid, Geyenna Sterling-Lentsch, Marc S. Halfon [DESCRIPTION] The collection, fixing, and immunohistochemical staining of Aedes aegypti embryos is challenging in comparison to D. melanogaster since the vitelline membrane of Ae. aegypti must b...
["[Preparation of an aspirator] Cut both ends from a 10 mL serological pipette. Save the end with the cotton plug.", "[Preparation of an aspirator] Trim the tapered tip and remove any sharp edges.", "[Preparation of an aspirator] Whittle the thinner plastic end of the serological pipette where the cotton plug is.", "[P...
88,224
10x Protocols: Chromium Single Cell/Nuclei Gene Expression Flex Multiplex -- University of Minnesota TMCs (CG000527 Rev D)
1
dx.doi.org/10.17504/protocols.io.bp2l6x81klqe/v1
https://www.protocols.io/view/10x-protocols-chromium-single-cell-nuclei-gene-exp-c2d8ya9w
IOx Genomics, Laura Niedernhofer
TITLE: 10x Protocols: Chromium Single Cell/Nuclei Gene Expression Flex Multiplex -- University of Minnesota TMCs (CG000527 Rev D) AUTHORS: IOx Genomics, Laura Niedernhofer [DESCRIPTION] 10x Genomics Chromium Single Cell Expression flex protocol for library construction. Protocol ID# (CG) and Revision letter provide...
["https://www.10xgenomics.com/products/single-cell-gene-expression-flex\nhttps://www.10xgenomics.com/support/single-cell-gene-expression-flex", "Complete single cell or nuclei isolation and 10x fixation prior to starting this protocol"]
25,317
RNA Isolation from Plant Tissue Protocol 11: Tri Reagent Method
null
dx.doi.org/10.17504/protocols.io.4ydgxs6
null
null
TITLE: RNA Isolation from Plant Tissue Protocol 11: Tri Reagent Method AUTHORS: [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Implemented by: Michael Melkonian and Barbara Surek</div><div class = "text-block"><span>The protocol for RNA isolation using </span><a href="https://www.protocols.io/view...
[]
36,723
Metabolomics and Lipidomics Sample Preparation
1
dx.doi.org/10.17504/protocols.io.bf4tjqwn
https://www.protocols.io/view/metabolomics-and-lipidomics-sample-preparation-bf4tjqwn
Kevin Contrepois
TITLE: Metabolomics and Lipidomics Sample Preparation AUTHORS: Kevin Contrepois [DESCRIPTION] Scope: To describe the procedure to extract metabolites and complex lipids using a biphasic separation with cold methyl tert-butyl ether (MTBE), methanol and water. Expected outcome/data: Metabolites and complex lipids are p...
["Roughly 30 mg of frozen tissues were homogenized in 500 µl ice-cold methanol by bead beating (MP bioscience cat# 6913-100, Solon, OH) at 4°C (2 x 45 s).", "1 ml of MTBE was added to 300 μl of the homogenate spiked-in with 40 µL deuterated lipid internal standards (Sciex, cat#: 5040156, lot#: LPISTDKIT-101).", "The sa...
17,415
Impact of glucose-6-phosphate dehydrogenase deficiency on dengue infection in Myanmar childrened protocol
null
dx.doi.org/10.17504/protocols.io.u9fez3n
null
Win Lai May, Myat Phone Kyaw, Stuart D. Blacksell, Sasithon Pukrittayakamee, Kesinee Chotivanich, Borimas Hanboonkunupakarn, Khin Nyo Thein, Chae Seung Lim, Janjira Thaipadungpanit, Thomas Althaus, podjanee jittamala
TITLE: Impact of glucose-6-phosphate dehydrogenase deficiency on dengue infection in Myanmar childrened protocol AUTHORS: Win Lai May, Myat Phone Kyaw, Stuart D. Blacksell, Sasithon Pukrittayakamee, Kesinee Chotivanich, Borimas Hanboonkunupakarn, Khin Nyo Thein, Chae Seung Lim, Janjira Thaipadungpanit, Thomas Althaus, ...
[]
64,458
XP Nutrition Keto Gummies: (Exposed 2022) Is XP Nutrition Keto Gummies Burn Fat? Where To Buy? Price!
4
dx.doi.org/10.17504/protocols.io.6qpvr6qjzvmk/v1
https://www.protocols.io/view/xp-nutrition-keto-gummies-exposed-2022-is-xp-nutri-ca7ishke
XP Nutrition Keto Gummies
TITLE: XP Nutrition Keto Gummies: (Exposed 2022) Is XP Nutrition Keto Gummies Burn Fat? Where To Buy? Price! AUTHORS: XP Nutrition Keto Gummies [DESCRIPTION] XP Nutrition Keto Gummies [STEPS] 1. ➢Product Name —XP Nutrition Keto Gummies ➢Main benefits —Helps to reduce stress, an...
["➢Product Name —XP Nutrition Keto Gummies\n➢Main benefits —Helps to reduce stress, anxiety, and depression\n➢Ingredients —CBD Gummies\n➢ Location —United States\n➢Dosage — As Prescribed on Bottle or Consu...
48,144
Identification chart for common Daphnia species in lakes in Michigan, and their most common parasites
3
null
https://www.protocols.io/view/identification-chart-for-common-daphnia-species-in-bs9qnh5w
Meghan Duffy, Kristel Sanchez
TITLE: Identification chart for common Daphnia species in lakes in Michigan, and their most common parasites AUTHORS: Meghan Duffy, Kristel Sanchez [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is a guide to the common Daphnia (& one Ceriodaphnia) species found in stratified lakes in Michigan...
[]
49,052
Pesticide Analysis Autotrace SOP
6
null
https://www.protocols.io/view/pesticide-analysis-autotrace-sop-bt54nq8w
Lara Frankson
TITLE: Pesticide Analysis Autotrace SOP AUTHORS: Lara Frankson [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">To test for neonics in turf runoff.</div></div> [STEPS] ?. [Prepare Autotrace for T120 Runoff Extractions (Neonics)] Turn on the system (doesn’t need to be turned off unless idle for >14 d...
["[Prepare Autotrace for T120 Runoff Extractions (Neonics)]\nTurn on the system (doesn’t need to be turned off unless idle for >14 days)Open the nitrogen valve on the tank (to the right of the -40C freezer).Make sure the pressure on the regulator is no more than 100 psi.", "[Prepare Autotrace for T120 Runoff Extraction...
61,972
Protocol for Dead-End Ultrafiltration using REXEED 25S for Isolation of Salmonella from Surface Water
4
dx.doi.org/10.17504/protocols.io.bp2l613p1vqe/v1
https://www.protocols.io/view/protocol-for-dead-end-ultrafiltration-using-rexeed-b8rurv6w
Autumn L Kraft, Manan Sharma, Jonathan G Frye, James E Wells, Abasiofiok Mark Ibekwe, NARMS EWG
TITLE: Protocol for Dead-End Ultrafiltration using REXEED 25S for Isolation of Salmonella from Surface Water AUTHORS: Autumn L Kraft, Manan Sharma, Jonathan G Frye, James E Wells, Abasiofiok Mark Ibekwe, NARMS EWG [DESCRIPTION] Protocol describes dead-end ultrafiltration (DEUF) using REXEED 25S for the recovery ...
["[Protocol] If needed, add control strain to water sample by gently tipping vial over the opening. Close sample bottle tightly and thoroughly mix sample before proceeding.", "[Protocol] Prepare for filtration by setting up Geotech pump in the biosafety cabinet (BSC). Ensure there is enough room around pump to include ...
95,789
Monosynaptic Rabies Tracing
4
dx.doi.org/10.17504/protocols.io.3byl4qp9jvo5/v1
https://www.protocols.io/view/monosynaptic-rabies-tracing-c9smz6c6
Alexandra Nelson, Michael Ryan
TITLE: Monosynaptic Rabies Tracing AUTHORS: Alexandra Nelson, Michael Ryan [DESCRIPTION] This protocol describes the steps for performing stereotaxic surgery in mice. It is applicable to intracranial injections (e.g. virus, drug) and placement of implants (e.g. optical fibers, electrode arrays) into targeted regions o...
["Rabies injections were performed in a BSL-2 surgical suite following the protocol described in dx.doi.org/10.17504/protocols.io.n2bvj6qynlk5/v1", "For modified, G-deleted rabies viruses, first inject a Cre-dependent helper virus (AAV-DIO-sTpEpB-GFP) to restrict expression of the EnvA receptor (TVA) and rabies glycopr...
91,042
Perforated patch electrophysiology recordings
4
dx.doi.org/10.17504/protocols.io.36wgq3y9olk5/v1
https://www.protocols.io/view/perforated-patch-electrophysiology-recordings-c46ayzae
enrico.zampese
TITLE: Perforated patch electrophysiology recordings AUTHORS: enrico.zampese [DESCRIPTION] In this protocol we detail the steps to perform ex-vivo brain slices perforated patch electrophysiology recordings. The protocol is originally meant to record from striatal MSNs, but can be adapted to other cell types. [BEFORE_...
["[Prepare patch pipettes] Turn on the Sutter P-1000 puller and enter the desired pull protocol.", "[Prepare patch pipettes] Insert a thick-walled borosilicate glass capillary and press pull.", "[Prepare patch pipettes] Pipette resistance must be of 3 to 6 megaohms.", "[Setting up patch rig and environment] Turn on the...
87,960
External Quality Control for inter-Batch comparison
1
dx.doi.org/10.17504/protocols.io.yxmvm391ol3p/v1
https://www.protocols.io/view/external-quality-control-for-inter-batch-compariso-cz5yx87w
Ricardo M. Borges
TITLE: External Quality Control for inter-Batch comparison AUTHORS: Ricardo M. Borges [DESCRIPTION] Controle de Qualidade Externo (QCExt): A amostra contendo a cepa código CCMR0280 cultivada em paralelo a todos os lotes servirá como referência para estudos comparativos estudo de comparação entre as cepas do CCMR (Cul...
["[Material] microtubos de 2 mL com tampa de rosca\ntudo de centrifugação de 50 mL tipo Falcon\nBalança analítica (AUW220 , Shimadzu)", "[Material em Estoque] Inicialmente, um lote inicial composto por 25 réplicas do cultivo da cepa código CCMR0280\nfoi (i) cultivado, (ii) coletado e combinado, (iii) submetido a liofil...
26,237
CULTURING i3LMNS (Basic Protocol 8)
null
dx.doi.org/10.17504/protocols.io.5u5g6y6
null
Michael S. Fernandopulle, Ryan Prestil, Christopher Grunseich, Chao Wang, Li Gan, Michael E. Ward
TITLE: CULTURING i3LMNS (Basic Protocol 8) AUTHORS: Michael S. Fernandopulle, Ryan Prestil, Christopher Grunseich, Chao Wang, Li Gan, Michael E. Ward [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Induction and differentiation of i</span><span style = "vertical-align:super;">3</span><span>LMN...
["[Coating dishes]\nPrepare stock solutions of PLO, PEI, or PDL (see Table 3).", "[Coating dishes]\nAdd one-half culture volume of 1× coating solution from step 1 to the tissue culture dishes to be used for plating Day 3 partially differentiated i3LMNs.", "[Coating dishes]\nIncubate dishes for at least at .\n0 Room te...
21,750
Copy of Immunohistochemistry
null
dx.doi.org/10.17504/protocols.io.zgwf3xe
null
Ning Zhang, Yu Kuang
TITLE: Copy of Immunohistochemistry AUTHORS: Ning Zhang, Yu Kuang [STEPS] ?. Deparaffinize/hydrate sections: a.Incubate sections in three washes of xylene for 5 minutes each.b.Incubate sections in two washes of 100% ethanol for 10 minutes each.c.Incubate sections in two washes of 95% ethanol for 10 minutes each. ?. Wa...
["Deparaffinize/hydrate sections: a.Incubate sections in three washes of xylene for 5 minutes each.b.Incubate sections in two washes of 100% ethanol for 10 minutes each.c.Incubate sections in two washes of 95% ethanol for 10 minutes each.", "Wash sections twice in dH2O for 5 minutes each.", "Incubate sections in 3% hyd...
28,865
Creatinine Clearance by HPLC
null
dx.doi.org/10.17504/protocols.io.8e9hth6
null
Kumar Sharma
TITLE: Creatinine Clearance by HPLC AUTHORS: Kumar Sharma [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">This protocol describes the procedures for collecting, preparing and performing the DiaComp Creatinine Clearance...
["[Collection of timed mouse urines.]\nGeneral procedure using the Nalgene Diuresis Cage (650-0322): a clean cage is prepared for use by lightly spraying the (1) metal mouse screen, (2) the mesh support grid and (3) the conical mouse funnel with silicone spray. Wipe off any excessive silicone and allow it to sit on ben...
106,631
PFF seeding in U2OS cells
0
dx.doi.org/10.17504/protocols.io.q26g717bqgwz/v1
https://www.protocols.io/view/pff-seeding-in-u2os-cells-dkdf4s3n
Isabel Lam, Alain Ndayisaba, Vikram Khurana
TITLE: PFF seeding in U2OS cells AUTHORS: Isabel Lam, Alain Ndayisaba, Vikram Khurana [DESCRIPTION] PFF seeding in U2OS cells [STEPS]
[]
33,325
Brooks Lab Western Blotting Protocol
null
dx.doi.org/10.17504/protocols.io.bcsmiwc6
null
Brooks Lab University of California, Eva Robinson, Alison Tang
TITLE: Brooks Lab Western Blotting Protocol AUTHORS: Brooks Lab University of California, Eva Robinson, Alison Tang [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is a Western Blotting Protocol for Brooks Lab, Department of Biomolecular Engineering, University of California, Santa Cruz.</div>...
["[RIPA Lysis Buffer]\nMake , cover, and freeze in 10 ml aliquots at . ORUse pre-made Pierce RIPA buffer, 100 ml, which is aliquoted in aliquots and stored in\n[RIPA w/o protease inhibitor]\n-20 °C\n10 ml\n-20 °C", "[RIPA Lysis Buffer]\nBefore use, thaw and add 1 tablet of protease inhibitor (PI) per 10 ml aliquot. (...
66,461
Purification and Crystallization of ATG9 HDIR-ATG101:ATG13 complex
4
dx.doi.org/10.17504/protocols.io.dm6gpbkdjlzp/v1
https://www.protocols.io/view/purification-and-crystallization-of-atg9-hdir-atg1-cc55sy86
Adam Yokom, Xuefeng Ren
TITLE: Purification and Crystallization of ATG9 HDIR-ATG101:ATG13 complex AUTHORS: Adam Yokom, Xuefeng Ren [DESCRIPTION] Purification and Crystallization of ATG9 HDIR (828-839) fused ATG101 (1-198):ATG13 (1-197) [STEPS] SECTION: Crystallization 19. ATG9 HDIR-ATG101:ATG13 complex at 6 mg/ml in 25 mM HEPES pH 7.5, 1...
["[Crystallization] ATG9 HDIR-ATG101:ATG13 complex at 6 mg/ml in 25 mM HEPES pH 7.5, 150mM NaCl, 1mM MgCl2, 1mM TCEP was used as the protein stock", "[Crystallization] Crystallization was carried out by sitting-drop vapor diffusion using an automated liquid-handling system (Mosquito, TTP LabTech, UK) at 288 K in 96-we...
15,198
SN medium
null
dx.doi.org/10.17504/protocols.io.s36egre
null
Roscoff Culture Collection
TITLE: SN medium AUTHORS: Roscoff Culture Collection [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Used for some cyanobacteria in particular </span><span style = "font-style:italic;">Synechococcus</span><span style = "font-style:italic;">.</span></div></div> [STEPS] ?. Autoclave and filter ...
["Autoclave and filter aged seawaterWork under laminar flow hood.Add to 800mL of seawater the following nutriments that have been autoclaved beforehand  (except vitamin) Quantity\n Compound\n Stock Solution\n 2,0 mL\n EDTA disodium salt dihydrate (Na2EDTA-2H2O)\n 2,8 g/L\n 2,0 mL\n Sodium carbonat...
null
null
null
dx.doi.org/10.17504/protocols.io.u73ezqn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This protocol results in the isolation of single nucleated zoospores from a culture of Aurantiochytrium limacinum. The point is to recover single nucleus colonies rather than single cell colonies (which may contain varying nuclei - particularly following transformation). [STEP...
["[Prepare Culture] Inoculate 500 ul GPY media (microcentrifuge tube) with toothpick scrape of a colony. If inoculating a zeocin resistant colony, add 5 ul zeocin (for 100 ug/ml). Vortex/ resuspend cells. Pipette 4.5 ml of GPY into a 15 ml glass culture tube. Add 500 ul inoculum. Incubate at 28 C, 170 rpm for 48 h, or ...
null
null
null
dx.doi.org/10.17504/protocols.io.ibbcain
null
null
TITLE: No Title AUTHORS: [BEFORE_START] <p>-Sterile polypropylene tubes (15 or 50 ml); sterile Oak Ridge-tubes</p> <p> </p> <p>-Steel forceps </p> <p> </p> <p>-Scissors</p> <p> </p> <p>-glass rod with rounded end</p> <p> </p> <p>-95% Ethanol for sterilization</p> <p> </p> <p>-Sterile lysis buffer (10 mM Tris, 100 mM ...
[]
51,686
Test protocol II
1
null
https://www.protocols.io/view/test-protocol-ii-bwqepdte
Abby Moore
TITLE: Test protocol II AUTHORS: Abby Moore [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is a test protocol</div></div> [STEPS] ?. Use 80:20 MeOH:H2O for this step. This is not easy to access by machine. ?. If you haven't already, make a solution with the following components: ?. If you hav...
["Use 80:20 MeOH:H2O for this step. This is not easy to access by machine.", "If you haven't already, make a solution with the following components:", "If you haven't already, make a solution with the following components:", "Use this piece of equipment:"]
71,137
Colony PCR
1
dx.doi.org/10.17504/protocols.io.dm6gpjwx8gzp/v1
https://www.protocols.io/view/colony-pcr-chp9t5r6
Georgina Elisa Diego Macías, Ana Belem García González, Irán Alessandra Chaparro Rodríguez, Jair Alexis Gardea Sáenz
TITLE: Colony PCR AUTHORS: Georgina Elisa Diego Macías, Ana Belem García González, Irán Alessandra Chaparro Rodríguez, Jair Alexis Gardea Sáenz [DESCRIPTION] Protocol for a PCR from a colony [STEPS] 1. Dissolve one colony in 20 µL of nuclease-free water and heat at 80°C for 10 minutes. 2. Prepare the following 50 uL...
["Dissolve one colony in 20 µL of nuclease-free water and heat at 80°C for 10 minutes.", "Prepare the following 50 uL reaction in a 0.2mL PCR tube on ice:", "Gently mix the reaction and spin down in the microcentrifuge", "Cycling conditions for a routine PCR:"]
26,190
DIFFERENTIATION OF i3LMNS (Basic Protocol 7)
null
dx.doi.org/10.17504/protocols.io.5tng6me
null
Michael S. Fernandopulle, Ryan Prestil, Christopher Grunseich, Chao Wang, Li Gan, Michael E. Ward
TITLE: DIFFERENTIATION OF i3LMNS (Basic Protocol 7) AUTHORS: Michael S. Fernandopulle, Ryan Prestil, Christopher Grunseich, Chao Wang, Li Gan, Michael E. Ward [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol describes the rapid and robust differentiation of hiPSCs into lower motor...
["[Day 0]\nPrepare a sufficient amount of hNIL-iPSC for differentiation.\nApproximately two wells of a 6-well dish at 80 % confluency are sufficient to begin the differentiation of a 10-cm dish, and cell numbers may be scaled by well surface area.Typically, hNIL lines proliferate for longer than hNGN2 lines, and this p...
9,378
Heterologous expression and affinity purification of Strep-tagged (KaiC) proteins
null
dx.doi.org/10.17504/protocols.io.meac3ae
null
Anika Wiegard, Christin Köbler, Katsuaki Oyama, Anja K. Dörrich, Chihiro Azai, Kazuki Terauchi, Annegret Wilde, Ilka Maria Axmann
TITLE: Heterologous expression and affinity purification of Strep-tagged (KaiC) proteins AUTHORS: Anika Wiegard, Christin Köbler, Katsuaki Oyama, Anja K. Dörrich, Chihiro Azai, Kazuki Terauchi, Annegret Wilde, Ilka Maria Axmann [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol can be use...
["[heterologous protein expression in E.coli]\ntransformation", "[heterologous protein expression in E.coli]\npre-culture:inoculate LB medium containing 75-100 µg ampicillin ml-1 (optional: plus 30 µg chloramphenicol ml-1) with resulting transformants (use e.g. 200 ml LB)incubate over night at 37 °C and 200-250 r.p.m."...
71,070
Nanopore sequencing data analysis using Microsoft Azure cloud computing service
5
dx.doi.org/10.17504/protocols.io.x54v9dj7pg3e/v1
https://www.protocols.io/view/nanopore-sequencing-data-analysis-using-microsoft-chm6t49e
Linh Truong
TITLE: Nanopore sequencing data analysis using Microsoft Azure cloud computing service AUTHORS: Linh Truong [DESCRIPTION] This protocol provides instruction to set up the analytic pipeline to process raw data from Oxford Nanopore Sequencing. This pipeline leverages the computing resources available in Microsoft Azure ...
["[Section 1: Generation of data on-site] Load the multiplexed HLA library pool consisting of 48 individuals onto a MinION flow cell. The data is acquired using MinKNOW software for 16 hours using default settings. \n \n960 min", "[Section 1: Generation of data on-site] The raw FAST5 files are stored in a local folder ...
null
null
null
dx.doi.org/10.17504/protocols.io.nupdevn
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?.
[]
27,302
Coulter Method for the detection of Physiological parameters
null
dx.doi.org/10.17504/protocols.io.6wehfbe
null
Xiaohua Du, Xia Liu, Mawolo James Blackar, Yingjie Zhou, Haifeng Wang, Fayang Liu, Zhiqing He
TITLE: Coulter Method for the detection of Physiological parameters AUTHORS: Xiaohua Du, Xia Liu, Mawolo James Blackar, Yingjie Zhou, Haifeng Wang, Fayang Liu, Zhiqing He [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Instrument</span></div><div class = "text-block...
[]
107,048
Astrocyte generation
0
dx.doi.org/10.17504/protocols.io.bp2l62e5dgqe/v1
https://www.protocols.io/view/astrocyte-generation-dksg4wbw
Patricia López García
TITLE: Astrocyte generation AUTHORS: Patricia López García [DESCRIPTION] Protocol to generate astrocytes from cortical NPCs using a comercial media (AM) [STEPS] SECTION: NPC generation 1. To prepare the iPSC’s for neural induction, iPSCs need to grow to 90 - 100% confluence. SECTION: NPC generation 2. As soon as the...
["[NPC generation] To prepare the iPSC’s for neural induction, iPSCs need to grow to 90 - 100% confluence.", "[NPC generation] As soon as the cells reach 90% confluence mTeSR media is switched to neural induction media (N2B27 supplemented with 10uM SB431542 and 1uM Dorsomorphin).", "[NPC generation] Neural induction me...
92,392
noesypr1d_metab.nan
5
dx.doi.org/10.17504/protocols.io.x54v9p21pg3e/v4
https://www.protocols.io/view/noesypr1d-metab-nan-c6ggzbtw
NAN KB, John Glushka, Mario Uchimiya, Saraa Al Jawad, Christopher Esselman, Leandro I Ponce, Laura Morris, Arthur Edison
TITLE: noesypr1d_metab.nan AUTHORS: NAN KB, John Glushka, Mario Uchimiya, Saraa Al Jawad, Christopher Esselman, Leandro I Ponce, Laura Morris, Arthur Edison [DESCRIPTION] This is a protocol for running the Bruker pulse program "noesypr1d". [BEFORE_START] This protocol assumes your sample is loaded, locked, tuned, and...
["[Create a new dataset]", "[Create a new dataset] On the menu bar on TopSpin, click on\nStart → Create Dataset", "[Create noesypr1d experiment file] When Lock, Tune, Shim are complete proceed to the specific protocol for your desired pulseprogram(s).", "[Create a new dataset] A new window opens. On the right top bar, ...
null
null
null
dx.doi.org/10.17504/protocols.io.grwbv7e
null
null
TITLE: No Title AUTHORS: [STEPS] ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.jqwcmxe
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?.
[]
44,420
Live Cell Quantification using Image Analysis
1
dx.doi.org/10.17504/protocols.io.bpmcmk2w
https://www.protocols.io/view/live-cell-quantification-using-image-analysis-bpmcmk2w
Minjung Song, Florian Merkle
TITLE: Live Cell Quantification using Image Analysis AUTHORS: Minjung Song, Florian Merkle [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Purpose</span></div><div class = "text-block">This protocol describes image-based quantification of human pluripotent stem cell...
["[Cell culture and staining: Cell plating]\nCoat plate with appropriate substrate to prepare cells for plating.\nWe have found the 24-well black μ-Plate from Ibidi allows good cell growth and is compatible with high-content imaging systems.", "[Cell culture and staining: Cell plating]\nWhen plating cells for culture, ...
69,505
PCR amplicon next generation sequencing
1
dx.doi.org/10.17504/protocols.io.36wgq64olk57/v3
https://www.protocols.io/view/pcr-amplicon-next-generation-sequencing-cf49tqz6
Susanne Kreutzer, Lilly van de Venn, Charles Yeh, Mark DeWitt
TITLE: PCR amplicon next generation sequencing AUTHORS: Susanne Kreutzer, Lilly van de Venn, Charles Yeh, Mark DeWitt [DESCRIPTION] Preparation of Amplicons for deep sequencing is based on two PCR steps. The genomic loci of interest is enriched in a first PCR including a stubber sequence to both ends of the molecule....
["[Extract genomic DNA from edited cells using QuickExtract solution] Resuspend cell pellet to ≥2,500 cells/µL in QuickExtract solution. Vortex or pipette-mix to resuspend thoroughly.\n \n\nMake sure to check whether QuickExtract is appropriate for your sample and application. Take a look at \n\nfor a more extensive li...
42,327
Unlocking Spatial Molecular &amp; Cellular Relationships of SARS-CoV-2 in Archived Human Tissue
3
dx.doi.org/10.17504/protocols.io.bmjxk4pn
https://www.protocols.io/view/unlocking-spatial-molecular-amp-cellular-relations-bmjxk4pn
Sam Pattle, Jenna Gregory, Fergal M Waldron
TITLE: Unlocking Spatial Molecular &amp; Cellular Relationships of SARS-CoV-2 in Archived Human Tissue AUTHORS: Sam Pattle, Jenna Gregory, Fergal M Waldron [DESCRIPTION] The true prevalence of SARS-CoV-2 infection in the UK population remains unknown. Phylogenetic diversity estimates derived from genomic sequence trac...
[]
42,342
Copy of Lab 1 Notebook
3
null
https://www.protocols.io/view/copy-of-lab-1-notebook-bmkek4te
Kamron Mojabe
TITLE: Copy of Lab 1 Notebook AUTHORS: Kamron Mojabe [STEPS]
[]
77,698
The monocyte-derived dendritic cell (MoDC) assay – an in vitro assay for testing the immunogenicity of cellular therapies
4
dx.doi.org/10.17504/protocols.io.8epv5jmqdl1b/v2
https://www.protocols.io/view/the-monocyte-derived-dendritic-cell-modc-assay-an-cp5avq2e
Annabel J Curle, Sarah Howlett, Joanne Jones
TITLE: The monocyte-derived dendritic cell (MoDC) assay – an in vitro assay for testing the immunogenicity of cellular therapies AUTHORS: Annabel J Curle, Sarah Howlett, Joanne Jones [DESCRIPTION] The monocyte-derived dendritic cell (MoDC) assay can be used to test, in vitro, the immunogenicity of potential regenerati...
["[Cell preparations and moDC differentiation - Day 0-7] Make PBMCs using preferred method from whole blood", "[Cell preparations and moDC differentiation - Day 0-7] Take whole PBMCs and enrich for CD14+ cells using magnetic activated cell sorting (MACS) with CD14+ microbeads", "[Cell preparations and moDC differentiat...
28,285
MojoSort™ Nanobeads Protocol - 1
null
dx.doi.org/10.17504/protocols.io.7u5hny6
null
Sam Li
TITLE: MojoSort™ Nanobeads Protocol - 1 AUTHORS: Sam Li [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Product description and procedure summary:</span></div><div class = "text-block">The cells targeted by the Nanobeads are either selected or depleted by incubating...
["Prepare cells from your tissue of interest or blood without lysing erythrocytes. Kits for human samples have been optimized for PBMCs, please prepare the cells using a suitable method.", "In the final wash of your sample preparation, resuspend the cells in MojoSort™ Buffer by adding up to 4mL in a 5 mL (12 x 75 mm) p...
26,381
Microbiome Assay
null
dx.doi.org/10.17504/protocols.io.5zmg746
null
Priota Islam
TITLE: Microbiome Assay AUTHORS: Priota Islam [STEPS] ?. Preparing worms Chunk N2s on 3-4 plates on Wednesday, also book the hood for next Wednesday morning to dry the seeded plates Bleach the N2s on Friday and keep the tube in the rotator inside the 20C incubator On Monday refeed the arrested L1s to be young adults o...
["Preparing worms Chunk N2s on 3-4 plates on Wednesday, also book the hood for next Wednesday morning to dry the seeded plates Bleach the N2s on Friday and keep the tube in the rotator inside the 20C incubator On Monday refeed the arrested L1s to be young adults on Thursday (Tracking Day1), do the same on Tuesday for t...
44,260
test 1 v3
1
null
https://www.protocols.io/view/test-1-v3-bpgcmjsw
611
TITLE: test 1 v3 AUTHORS: 611 [STEPS] ?. test
["test"]
null
null
null
dx.doi.org/10.17504/protocols.io.dzr755
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] The following continued adaptation of Hennes et al. (1995) is a result of FLVP assay optimizations for the <em>Vibrio alginolyticus</em> PWH3a-phage P1 phage-host system (PHS) for use with SYBR Green I stain. Novices to microscopy would benefit from reading Wen et al. (2004) for...
[]
87,957
A computational pipeline to quantify perinuclear lysosomes in fibroblasts using CellProfiler
5
dx.doi.org/10.17504/protocols.io.81wgbxrw3lpk/v1
https://www.protocols.io/view/a-computational-pipeline-to-quantify-perinuclear-l-cz5vx866
Ebsy Jaimon, Suzanne R Pfeffer
TITLE: A computational pipeline to quantify perinuclear lysosomes in fibroblasts using CellProfiler AUTHORS: Ebsy Jaimon, Suzanne R Pfeffer [DESCRIPTION] Here we present a CellProfiler software pipeline to quantify the distribution of lysosomes in MEF cells. The lysosomes were stained using anti-LAMP1 antibody, and nu...
["[Import files into CellProfiler and extract metadata] A. Select the Images module, drag and drop the maximum intensity projected .TIF files as indicated\n\nB. Select the Metadata module\n\nIn the Metadata module: \n\nExtract metadata? Yes.\n\nMetadata extraction method: Extract from file/folder names\n\nMetadata so...
93,094
Culture of human epithelial cells (skin, cornea, thymus) on 3T3J2 feeder layer cells
4
dx.doi.org/10.17504/protocols.io.14egn31ezl5d/v3
https://www.protocols.io/view/culture-of-human-epithelial-cells-skin-cornea-thym-c66ezhbe
Pierluigi Giuseppe Manti
TITLE: Culture of human epithelial cells (skin, cornea, thymus) on 3T3J2 feeder layer cells AUTHORS: Pierluigi Giuseppe Manti [DESCRIPTION] Despite several protocols having been published in the past in a fragmented way, this protocol represents a harmonious recollection and reorganization of those, which proves to be...
["[3T3J2 Growth Medium] Open DMEM bottle (ThermoFisher Scientific).", "[3T3J2 Growth Medium] Add 8 % (v/v) of bovine serum (ThermoFisher Scientific) (DO NOT use fetal calf serum!).", "Set up one flask at low density to maintain the line (5*10^5 cells at early passages, 10^5 cells at late passages). This flask should r...
74,817
ARTIC-like Bacillus anthracis MLVA amplicon sequencing protocol for MinION
4
dx.doi.org/10.17504/protocols.io.3byl4jzozlo5/v1
https://www.protocols.io/view/artic-like-bacillus-anthracis-mlva-amplicon-sequen-cma9u2h6
Ágnes Nagy, Gábor Tóth
TITLE: ARTIC-like Bacillus anthracis MLVA amplicon sequencing protocol for MinION AUTHORS: Ágnes Nagy, Gábor Tóth [DESCRIPTION] Multiple-Locus Variable Number of Tandem Repeats (VNTR) Analysis (MLVA) is one of the gold standard strain-level subtyping methods for outbreak-related Bacillus anthracis strains. The repeat...
["[Primer pool preparation] If required resuspend lyophilised primers at a concentration of 100µM each. Primer names, characteristics, concentrations and volumes required for primer stocks are listed in the table below.", "[Primer pool preparation] Generate 500 µL primer Pool 1 stock by adding 7 µL, 13.5 µL or 15.5 µL ...
41,873
Fluorescent-Reporter Based Assay
4
dx.doi.org/10.17504/protocols.io.bk5rky56
https://www.protocols.io/view/fluorescent-reporter-based-assay-bk5rky56
Piyush Jain, Long T Nguyen, Santosh Rananaware
TITLE: Fluorescent-Reporter Based Assay AUTHORS: Piyush Jain, Long T Nguyen, Santosh Rananaware [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The CRISPR-Enhance SARS-CoV-2 detection kit has been designed to detect fragments of the Nucleocapsid (“N”) gene and Envelope gene (E) of SARS-CoV-2. An in...
["[RT-LAMP Master Mix Preparation]\nLabel a new microcentrifuge tube for each target ( N, E and RP) and prepare a RT-LAMP Master Mix consisting of the WarmStart®Colorimetric LAMP 2X Master Mix with UDG.and the appropriate 10x Primer Mix using the recipe in Table 1 below. Make enough of each master mix for all samples ...
43,254
BGISEQ-500 WGS library construction
1
null
https://www.protocols.io/view/bgiseq-500-wgs-library-construction-bngwmbxe
Jie Huang, Xinming Liang, Yuankai Xuan, Chunyu Geng, Yuxiang Li, Haorong Lu, Shoufang Qu, Xianglin Mei, Hongbo Chen, Ting Yu, Nan Sun, Junhua Rao, Jiahao Wang, Wenwei Zhang, Ying Chen, Sha Liao, Hui Jiang, Xin Liu, Zhaopeng Yang, Feng Mu, Shangxian Gao
TITLE: BGISEQ-500 WGS library construction AUTHORS: Jie Huang, Xinming Liang, Yuankai Xuan, Chunyu Geng, Yuxiang Li, Haorong Lu, Shoufang Qu, Xianglin Mei, Hongbo Chen, Ting Yu, Nan Sun, Junhua Rao, Jiahao Wang, Wenwei Zhang, Ying Chen, Sha Liao, Hui Jiang, Xin Liu, Zhaopeng Yang, Feng Mu, Shangxian Gao [DESCRIPTION] ...
["[Overview]", "[DNA fragmentation]\n1) Input genomic DNA sampleGenomic DNA Sample Recommendation AB1Nucleic Acid\n*High-quality genomic DNA\n2Molecular Weight\n&gt;23k bp\n3Amount\n1μg\n4Concentration\n≥12.5ng/μL\n5Purity\nOD260/OD280=1.8~2.0\n*High-quality genomic DNA should be free of salt or organics. It could run...
86,194
TMT labelling
4
null
https://www.protocols.io/view/tmt-labelling-cyesxtee
Louise Uoselis
TITLE: TMT labelling AUTHORS: Louise Uoselis [DESCRIPTION] TMT labelling [STEPS] 1. Reconstitute lyophilised peptides in 25 µL by vortexing each sample for ~10 s, and then leaving to sit at Room temperature for 15 min. Each sample was then sonicated for 5 min in a waterbath sonicator in an ice slurry. 2. Determine ...
["Reconstitute lyophilised peptides in 25 µL by vortexing each sample for ~10 s, and then leaving to sit at Room temperature for 15 min. Each sample was then sonicated for 5 min in a waterbath sonicator in an ice slurry.", "Determine the peptide concentration of each sample spectroscopically", "Aliquot10 µg of peptides...
61,884
Immunostaining of Bodo saltans
4
null
https://www.protocols.io/view/immunostaining-of-bodo-saltans-b8n4rvgw
Ewa Chrostek, Mastaneh Ahrar, Gregory Dd Hurst
TITLE: Immunostaining of Bodo saltans AUTHORS: Ewa Chrostek, Mastaneh Ahrar, Gregory Dd Hurst [DESCRIPTION] This protocol is used in our Laboratory in Liverpool to perform IF on Bodo saltans cells. [STEPS] SECTION: Culture conditions 1. Bodo saltans was cultured in a cerophyl-based medium enriched with 3.5 mM s...
["[Culture conditions] Bodo saltans was cultured in a cerophyl-based medium enriched with 3.5 mM sodium phosphate dibasic (Na2HPO4)1. Cultures were incubated at 22 °C in T25 tissue culture flasks containing 20 ml of media bacterized with Klebsiella pneumoniae subsp. Pneumoniae (ATCC® 700831).", "[Immunostaining] Please...
43,379
A Simple Multistep Protocol for Differentiating Human Induced Pluripotent Stem Cells into Functional Macrophages
1
dx.doi.org/10.17504/protocols.io.bnktmcwn
https://www.protocols.io/view/a-simple-multistep-protocol-for-differentiating-hu-bnktmcwn
Chandrayana Mukherjee, Christine Hale, Subhankar Mukhopadhyay
TITLE: A Simple Multistep Protocol for Differentiating Human Induced Pluripotent Stem Cells into Functional Macrophages AUTHORS: Chandrayana Mukherjee, Christine Hale, Subhankar Mukhopadhyay [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Macrophages differentiated from human induced pluripotent ste...
["[General Considerations]\nHuman iPSCs are delicate and more difficult to maintain in culture compared to most other conventional human cell lines. They show a natural tendency to spontaneously differentiate into fibroblast like cells. Cytokines and growth factors present in the medium allow them to be kept in pluripo...
71,010
Participant Registration Form For Mapping the Early Childhood Gut Across Ancestry, Geography, and Environment (Gut-AGE)
1
dx.doi.org/10.17504/protocols.io.6qpvr4d3zgmk/v1
https://www.protocols.io/view/participant-registration-form-for-mapping-the-earl-chkat4se
Fatima Zulqarnain, Stephanie Regis, elsy.ngwa, Asra Usmani, jason.boisvert, Arg7Ef, Sana Syed, Jocelyn Silvester, jay.thiagarajah
TITLE: Participant Registration Form For Mapping the Early Childhood Gut Across Ancestry, Geography, and Environment (Gut-AGE) AUTHORS: Fatima Zulqarnain, Stephanie Regis, elsy.ngwa, Asra Usmani, jason.boisvert, Arg7Ef, Sana Syed, Jocelyn Silvester, jay.thiagarajah [DESCRIPTION] The gastrointestinal (GI) system has a...
["[To be filled out by the study team] PARTICIPANT REGISTRATION FORM \n\nThe first part of the form collects registration information from each participant. The purpose of this section is to ensure that the study team has all relevant participant information for accessing participant health records at their site of car...
77,406
HYPROP soil sample collection
1
dx.doi.org/10.17504/protocols.io.261ge3jedl47/v1
https://www.protocols.io/view/hyprop-soil-sample-collection-cpt6vnre
marshall.bennett
TITLE: HYPROP soil sample collection AUTHORS: marshall.bennett [DESCRIPTION] Procedure developed using information gathered from the following sources: Shokrana, M. S. B., & Ghane, E. (2020, February 22). Measurement of soil water characteristic curve using hyprop2. Measurement of soil water characteristic curve usi...
["Gather tools needed for HYPROP sample ring collection.", "Identify sampling locations. Collect and record all appropriate field data (GPS coordinates, soil texture information, weather, etc.).", "Clear the surface of the soil to remove excess plant cover or debris. DO NOT DISTURB THE SOIL It is important to note that...
75,388
Insert Alignment ELYRA7
1
null
https://www.protocols.io/view/insert-alignment-elyra7-cmu4u6yw
Rafael Camacho
TITLE: Insert Alignment ELYRA7 AUTHORS: Rafael Camacho [DESCRIPTION] Alignment of the stage insert to ensure that the sample is perpendicular to the optical axis. This protocol is based on material provided by ZEISS' application specialists. What is needed before to start: - This protocol was designed for the EL...
["[Find sample plane via the eye pieces] ZEN black -> Locate Tab,\n \nand set an imaging protocol for transmitted light so you can see the sample via the eye pieces.", "[Find sample plane via the eye pieces] Change the objective to the lowest magnification, typically 10X", "[Find sample plane via the eye pieces] Place...
83,281
Sectioning of Mouse Brain by Microtome
1
dx.doi.org/10.17504/protocols.io.81wgbxd91lpk/v1
https://www.protocols.click/view/sectioning-of-mouse-brain-by-microtome-cvjrw4m6
Maryana Nissan
TITLE: Sectioning of Mouse Brain by Microtome AUTHORS: Maryana Nissan [DESCRIPTION] This protocol describes how to use the microtome to prepare and slice mouse brain sections for Immunohistochemistry [STEPS] SECTION: Preparation Methods 1. While preparing the microtome, make sure the blade is covered. The microtome ...
["[Preparation Methods] While preparing the microtome, make sure the blade is covered. The microtome used in the lab is from Leica Microsystems.", "[Preparation Methods] Slide the plate into the stage and secure it by tightening it with a screw.", "[Preparation Methods] Orient the plate so that it is centered.", "[Prep...
99,811
4% PFA + 2.5% Glutaraldehyde Fixative
1
dx.doi.org/10.17504/protocols.io.dm6gpr15dvzp/v3
https://www.protocols.io/view/4-pfa-2-5-glutaraldehyde-fixative-ddqb25sn
Allen Institute for Brain Science
TITLE: 4% PFA + 2.5% Glutaraldehyde Fixative AUTHORS: Allen Institute for Brain Science [DESCRIPTION] This protocol is used to prepare 4% Paraformaldehyde (PFA) + 2.5% Glutaraldehyde Fixative. This is utilized to fix the tissue after electrophysiology recordings. Note: Research reported in this publication was suppo...
[]
21,563
UC Davis - Vertical Sleeve Gastrectomy
null
dx.doi.org/10.17504/protocols.io.za3f2gn
null
Kristin Evans
TITLE: UC Davis - Vertical Sleeve Gastrectomy AUTHORS: Kristin Evans [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span></div><div class = "text-block"> Vertical sleeve Gastrectomy (VSG) is a common surgical treatment of obesity, type-2 diabetes, and oth...
["Preparation: 1. Animals should be fasted overnight to ensure that their stomach and intestines are as empty as possible. 2. Weight prior to surgery to calculate drugs (Meloxicam and Baytril dosing) 3. Autoclave all supplies and tools needed for the surgery, start the bead sterilizer for sterilizing instruments. 4. Pr...
108,247
Collection of Selected Behavioural Assessment Methods in the Tropical Marine Amphipod, Parhyale hawaiensis 
0
dx.doi.org/10.17504/protocols.io.eq2lywrowvx9/v1
https://www.protocols.io/view/collection-of-selected-behavioural-assessment-meth-dmxx47pn
Ibrahim Lawan
TITLE: Collection of Selected Behavioural Assessment Methods in the Tropical Marine Amphipod, Parhyale hawaiensis  AUTHORS: Ibrahim Lawan [DESCRIPTION] This is a collection of some selected protocols for Behavioural Assessment Methods in the Tropical Marine Amphipod, Parhyale hawaiensis  [STEPS]
[]
82,483
U54 SCENT Normal Lung Surgical Tissue Collection Procedure
1
null
https://www.protocols.click/view/u54-scent-normal-lung-surgical-tissue-collection-p-custwwen
Carolyn Glass, andrew.nixon, Matthew Hartwig, Nancy McGreal, SSCRS, Mary Jordan*
TITLE: U54 SCENT Normal Lung Surgical Tissue Collection Procedure AUTHORS: Carolyn Glass, andrew.nixon, Matthew Hartwig, Nancy McGreal, SSCRS, Mary Jordan* [DESCRIPTION] This document outlines the required criteria for inclusion and collection of lung tissue obtained by surgical resection at Duke University Hospital t...
["[Inclusion and Exclusion Criteria] Inclusion Criteria: \n • Any age \n • All sexes \n • Grossly normal lung obtainable through surgical pathology\n\nExclusion Criteria:\n • No recent treatment with chemotherapy (recent <1 year) \n • No recent treatment with radiation (recent <1 year) \n • No history...
21,615
CUT&RUN: Targeted in situ genome-wide profiling with high efficiency for low cell numbers
null
dx.doi.org/10.17504/protocols.io.zcpf2vn
null
Derek Janssens, Steven Henikoff
TITLE: CUT&RUN: Targeted in situ genome-wide profiling with high efficiency for low cell numbers AUTHORS: Derek Janssens, Steven Henikoff [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>We previously described a novel alternative to Chromatin Immunoprecipitation, Cleavage Under Targets & Relea...
["[Binding cells to beads (~30 min)]\nHarvest fresh culture(s) at room temperature and count cells. The same protocol can be used for up to 500,000 mammalian cells per sample and/or digestion time point.\nCRITICAL STEP: All steps prior to the addition of antibody are performed at room temperature to minimize stress on ...
103,197
Immunohistochemistry on mouse brain sections
0
dx.doi.org/10.17504/protocols.io.j8nlk8m4xl5r/v1
https://www.protocols.io/view/immunohistochemistry-on-mouse-brain-sections-dgz53x86
Shiyi Wang
TITLE: Immunohistochemistry on mouse brain sections AUTHORS: Shiyi Wang [DESCRIPTION] Immunohistochemistry on mouse brain sections [STEPS] 1. **Anesthesia and Perfusion** 1.1. - Anesthetize mice with 200 mg/kg Avertin. 1.2. - Perform transcardial perfusion using TBS/Heparin followed by 4% paraformaldehyde (PFA). 2. *...
["**Anesthesia and Perfusion**", "- Anesthetize mice with 200 mg/kg Avertin.", "- Perform transcardial perfusion using TBS/Heparin followed by 4% paraformaldehyde (PFA).", "**Brain Collection and Post-Fixation**", "- Remove the brain and post-fix it in 4% PFA overnight at 4°C.", "**Cryoprotection and Storage**", "- Cry...
50,273
Coral Carbohydrate Assay for 96-well plates
1
dx.doi.org/10.17504/protocols.io.bvb9n2r6
https://www.protocols.io/view/coral-carbohydrate-assay-for-96-well-plates-bvb9n2r6
Colleen Bove, Justin Baumann
TITLE: Coral Carbohydrate Assay for 96-well plates AUTHORS: Colleen Bove, Justin Baumann [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><ol style = "list-style-type: decimal;"></ol></div><div class = "text-block">1. Masuko T, Minami A, Iwasaki N, Majima T, Nishimura S, Lee YC. Carbohydrate analysis...
["[Carbohydrate Assay]\nPull desired coral slurries from the -80 freezer to thaw at room temperature", "[Carbohydrate Assay]\nCollect test tubes and label (10 for the standard + the number of samples running)", "[Carbohydrate Assay]\nMake the standards and blank as shown below Tube IDConcentration (mg/mL)Vol water (uL)...
45,087
Quantitative Assessment of Islet Viability upon Arrival to HIPP by Staining with Trypan Blue Dye
4
dx.doi.org/10.17504/protocols.io.bp97mr9n
https://www.protocols.io/view/quantitative-assessment-of-islet-viability-upon-ar-bp97mr9n
IIDP-HIPP
TITLE: Quantitative Assessment of Islet Viability upon Arrival to HIPP by Staining with Trypan Blue Dye AUTHORS: IIDP-HIPP [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;font-style:italic;">This Standard Operating Procedure (SOP) is based on the Vanderbilt Human Isl...
["[Procedures]\nDispersion of islets into single cell suspension", "[Procedures]\nPrepare 2 aliquots of approximately 100 handpicked islets in tubes to determine viability of islets. Note: This is important especially for islet preparations with lower purity where cell death in the contaminating exocrine compartment ma...
32,975
3.7% Paraformaldehyde solution
null
dx.doi.org/10.17504/protocols.io.bcfpitmn
null
Institute of Medical Biotechnology
TITLE: 3.7% Paraformaldehyde solution AUTHORS: Institute of Medical Biotechnology [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Protocol for preparing a 3.7% Paraformaldehyde (PFA) solution. 3.7% PFA solution can be used to fix tissues. </div></div> [STEPS] ?. For , add to a glass beaker on a st...
["For , add to a glass beaker on a stir plate.\n[3.7% Formaldehyde]\n[1X DPBS]\nBe sure to work under a ventilated fume hood.", "Heat while stirring to approximately .\n55 °C", "Add to the heated DPBS solution.\n[paraformaldehyde powder]", "Keep stirring until all PFA powder is dissolved.\nTake care that the solution...
null
null
null
dx.doi.org/10.17504/protocols.io.h4kb8uw
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?.
[]
21,241
Impact of preventive oral health interventions during the prenatal care on preterm delivery, an integrative review protocol.
null
dx.doi.org/10.17504/protocols.io.yyzfxx6
null
Cristina Dutra Vieira, Andreza Nayla de Assis Aguiar, Camila Aparecida Silva de Oliveira Lima, Zilma Reis
TITLE: Impact of preventive oral health interventions during the prenatal care on preterm delivery, an integrative review protocol. AUTHORS: Cristina Dutra Vieira, Andreza Nayla de Assis Aguiar, Camila Aparecida Silva de Oliveira Lima, Zilma Reis [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Backg...
["Search strategyThe search strategy was performed using the following keyword combination for the composition of key-words and descriptors of health: Health Education; Health Education, Dental; Health Promotion; Pregnant Women; Pregnancy Outcome; Infant, Low Birth Weight; Premature Birth; Infant, Small for Gestational...
48,666
Genome-wide quantification of TF binding at single DNA molecule resolution
1
dx.doi.org/10.17504/protocols.io.btr2nm8e
https://www.protocols.io/view/genome-wide-quantification-of-tf-binding-at-single-btr2nm8e
Rozemarijn Kleinendorst, guido.barzaghi , Mike L. Smith, Judith B. Zaugg, Arnaud Krebs
TITLE: Genome-wide quantification of TF binding at single DNA molecule resolution AUTHORS: Rozemarijn Kleinendorst, guido.barzaghi , Mike L. Smith, Judith B. Zaugg, Arnaud Krebs [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Precise control of gene expression requires the coordinated act...
[]
91,933
HyDrop-RNA v1.0
1
dx.doi.org/10.17504/protocols.io.dm6gpwqjjlzp/v3
https://www.protocols.io/view/hydrop-rna-v1-0-c5z5y786
Florian De Rop, Suresh Poovathingal, Stein Aerts
TITLE: HyDrop-RNA v1.0 AUTHORS: Florian De Rop, Suresh Poovathingal, Stein Aerts [DESCRIPTION] Step-by-step protocol for performing HyDrop-RNA. The duration of each step assumes an experienced protocol user. For a first-time user, we recommend doubling the expected time for each step. [STEPS] SECTION: Microfluidics P...
["[Microfluidics Preparation] Setting up the Microfluidic Framework\nThese steps need to happen in advance before the run can be started, as time between the preparation of the cell resuspension in RT mix and actual encapsulation needs to be minimised to preserve cell viability.", "[Microfluidics Preparation] Boot up ...
47,189
Protocol for assembling SARS-Cov-2 runs with s-aligner
5
dx.doi.org/10.17504/protocols.io.bsbvnan6
https://www.protocols.io/view/protocol-for-assembling-sars-cov-2-runs-with-s-ali-bsbvnan6
Juanjo Bermúdez
TITLE: Protocol for assembling SARS-Cov-2 runs with s-aligner AUTHORS: Juanjo Bermúdez [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is a protocol for using Contignant s-aligner to de-novo assemble SARS-Cov-2 genomes. S-aligner outperforms common open-source software for de novo-assembly of ...
["[Indexing and fast-testing]", "[Indexing and fast-testing]\nresults/your-run-id-500.fa should have the largest contig bigger than 1.200bp and at least one of the files containing aligned reads to form a contig should look like that:", "[Indexing and fast-testing]", "Trim your data and start again.", "[Indexing and fa...
98,706
RNA extraction using the PureLink® RNA Mini Kit
0
dx.doi.org/10.17504/protocols.io.261ge54r7g47/v1
https://www.protocols.io/view/rna-extraction-using-the-purelink-rna-mini-kit-dcms2u6e
Enrico Bagnoli, Miratul Muqit
TITLE: RNA extraction using the PureLink® RNA Mini Kit AUTHORS: Enrico Bagnoli, Miratul Muqit [DESCRIPTION] This Protocol details the extraction of total RNA using the PureLink® RNA Mini Kit. [STEPS] SECTION: Prepare lysates from ≤5 × 106 monolayer cells: 1. Remove the growth medium from the cells. SECTION: Prep...
["[Prepare lysates from ≤5 × 106 monolayer cells:] Remove the growth medium from the cells.", "[Prepare lysates from ≤5 × 106 monolayer cells:] Using RNase-free pipette tips, add the appropriate volume of lysis buffer prepared with 2-mercaptoethanol to sample.", "[Prepare lysates from ≤5 × 106 monolayer cells:] Proceed...
25,239
RNA Isolation from Plant Tissue Protocol 1: Qiagen RNeasy Plant Mini Kit
null
dx.doi.org/10.17504/protocols.io.4vxgw7n
null
Jim Leebens-Mack, Charlotte Carrigan
TITLE: RNA Isolation from Plant Tissue Protocol 1: Qiagen RNeasy Plant Mini Kit AUTHORS: Jim Leebens-Mack, Charlotte Carrigan [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Protocol implemented by: Jim Leebens-Mack and Charlotte Carrigan </div><div class = "text-block"><span>The first protocol used...
[]
59,959
Detection of recombinant and endogenous LPPR3 by western blot
1
dx.doi.org/10.17504/protocols.io.81wgb6z2olpk/v2
https://www.protocols.io/view/detection-of-recombinant-and-endogenous-lppr3-by-w-b6sxrefn
cristina.kroon
TITLE: Detection of recombinant and endogenous LPPR3 by western blot AUTHORS: cristina.kroon [DESCRIPTION] This is a protocol for detection of overexpressed and endogenous LPPR3 from N1E-115 cells and primary hippocampal neurons. [STEPS] SECTION: Sample preparation 1. N1E cells for detection of recombinant LPPR3 ...
["[Sample preparation] N1E cells for detection of recombinant LPPR3\n\nDIV 0: N1E cells were plated at a density of 150 000 cells/well (6-well plates) and grown overnight in DMEM medium (Gibco) with 10% FCS and 1% penicillin/streptomycin. \nDIV 1: The cells were transfected with 1 µg of DNA using Lipofectamine 2000 (Th...
82,749
Live imaging and analysis to investigate phase separation properties of NEMO during mitophagy
1
dx.doi.org/10.17504/protocols.io.j8nlkw7rxl5r/v1
https://www.protocols.io/view/live-imaging-and-analysis-to-investigate-phase-sep-cu25wyg6
OLIVIA HARDING, Erika L.F. Holzbaur
TITLE: Live imaging and analysis to investigate phase separation properties of NEMO during mitophagy AUTHORS: OLIVIA HARDING, Erika L.F. Holzbaur [DESCRIPTION] Phase condensation of biomolecular particles is increasingly examined for its relevance in physiological contexts. When we observed formation of NEMO puncta as...
["[Replace Standard Media with Imaging Media and Induce Mitophagy] Wait until imaging chamber is heated to 37 °C.", "[Replace Standard Media with Imaging Media and Induce Mitophagy] Aspirate media from sample.", "[Replace Standard Media with Imaging Media and Induce Mitophagy] Repeat the following two steps for a total...
null
null
null
dx.doi.org/10.17504/protocols.io.scneave
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol outlines our preparation of single-nuclei suspension from surgically acquired fresh human adult brain tissue.</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
33,743
MicroRNA Affymetrix microarray probe labeling from ocular fluids
1
dx.doi.org/10.17504/protocols.io.bc7pizmn
https://www.protocols.io/view/microrna-affymetrix-microarray-probe-labeling-from-bc7pizmn
Zeljka Smit-Mcbride
TITLE: MicroRNA Affymetrix microarray probe labeling from ocular fluids AUTHORS: Zeljka Smit-Mcbride [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Sample collection, isolation of miRNAs, and quality control </span></div><div class = "text-block"><span>Aqueous and ...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.pxidpke
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This is the protocol for the PhireKit touchdown PCR assay</p> [BEFORE_START] <p>The assay has been optimized and validated for the Bio-rad C1000 Touch Thermocycler.</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.f9tbr6n
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Protocol modified for <em>Puccinia striiformis</em> f. sp <em>tritici</em> (<em>Pst</em>), based on:</p> <p>Michielse, C. B., Hooykaas, P. J., van den Hondel, C. A., &amp; Ram, A. F. (2008). Agrobacterium-mediated transformation of the filamentous fungus Aspergillus awamori. ...
[]
69,101
Roadmap to the bioinformatic study of gene and protein phylogeny and evolution - a practical guide
5
dx.doi.org/10.17504/protocols.io.36wgq77e3vk5/v2
https://www.protocols.io/view/roadmap-to-the-bioinformatic-study-of-gene-and-pro-cfqmtmu6
florian.jacques, Paulina Bolivar, Kristian Pietras, Emma Hammarlund
TITLE: Roadmap to the bioinformatic study of gene and protein phylogeny and evolution - a practical guide AUTHORS: florian.jacques, Paulina Bolivar, Kristian Pietras, Emma Hammarlund [DESCRIPTION] We present a compilation of nucleic acid and protein databases and bioinformatic tools for phylogenetic reconstructions a...
["[Phylogenetic analysis] Option 1: Maximum parsimony\n\nMaximum parsimony is a classic and simple method, that calculates the minimum number of evolutionary steps, including nucleotide insertions, deletions or substitutions, between species.\n\nHowever, this method ignores hidden mutations and does not take into accou...
76,690
S. O. C. medium
4
null
https://www.protocols.io/view/s-o-c-medium-cn5svg6e
Andreas Sagen
TITLE: S. O. C. medium AUTHORS: Andreas Sagen [DESCRIPTION] S.O.C. Medium is used in the final step of bacterial cell transformation to obtain maximal transformation efficiency ofE. coli. S.O.C. [STEPS] SECTION: 500 mL 1 M Glucose solution 1. Fill the bottle with 400 mL distilled water SECTION: 500 mL 1 M Glucose sol...
["[500 mL 1 M Glucose solution] Fill the bottle with 400 mL distilled water", "[500 mL 1 M Glucose solution] Measure 90.08 g Glucose\n\nMaterials:", "[500 mL 1 M Glucose solution] Add distilled water to a total of 500 mL", "[500 mL 1 M Glucose solution] Filter sterilize with a 0.2 µm filter", "[1 000 mL S. O. B. medium...
41,088
Doctor Vida Pocket - COVID-19 assay - Instructions for Use
4
dx.doi.org/10.17504/protocols.io.bkc8kszw
https://www.protocols.io/view/doctor-vida-pocket-covid-19-assay-instructions-for-bkc8kszw
goncalo.doria
TITLE: Doctor Vida Pocket - COVID-19 assay - Instructions for Use AUTHORS: goncalo.doria [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Instruction of use for Doctor Vida pocket - COVID-19 assay</div></div> [STEPS] ?. [Preparation setup] Turn on Dr. Vida Pocket device - by plugging the Dr. Vida P...
["[Preparation setup]\nTurn on Dr. Vida Pocket device - by plugging the Dr. Vida Pocket device's power supply into a suitable power plug (120V/240V AC) and connecting the micro-usb plug into the Dr. Vida Pocket device.\nWhen turned on, the device will emit a short bip.", "[Start new assay]\nPress \"Start new assay\" in...
null
null
null
dx.doi.org/10.17504/protocols.io.qz7dx9n
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
91,120
CAR T cell characterization by flow cytometry
4
null
https://www.protocols.io/view/car-t-cell-characterization-by-flow-cytometry-c48qyzvw
Andrea R Daniel
TITLE: CAR T cell characterization by flow cytometry AUTHORS: Andrea R Daniel [DESCRIPTION] Flow cytometry of tumor tissue and blood from mice with human tumors. Mice bearing HCC1954 tumors were euthanized at days 3 and 19 post CAR T cel delivery. This protocol describes methods for preparing input CAR T cells and tum...
["[Tissue collection] Place mice under deep isoflurane anesthesia (5% isoflurane, minimum of 15 minutes)", "[Tissue collection] Spray the abdomen of the mouse with 70% ethanol, this is to avoid fur contamination of samples", "[Tissue collection] Perform a bilateral thoracotomy, cut the right atrium of the heart.", "[Ti...
55,929
Manual Nanotrap Concentration and RNA Extraction for SARS-CoV-2 Viral Capture
4
dx.doi.org/10.17504/protocols.io.b2uzqex6
https://www.protocols.io/view/manual-nanotrap-concentration-and-rna-extraction-f-b2uzqex6
Matthew Cavallo, Stephen P Hilton, Jamie VanTassell, Julia Raymond, Marlene K Wolfe, Pengbo Liu, Christine Moe
TITLE: Manual Nanotrap Concentration and RNA Extraction for SARS-CoV-2 Viral Capture AUTHORS: Matthew Cavallo, Stephen P Hilton, Jamie VanTassell, Julia Raymond, Marlene K Wolfe, Pengbo Liu, Christine Moe [DESCRIPTION] This protocol details a method for SARS-CoV-2 capture and concentration through the use of Na...
["[Concentration Procedure] Place a 50 mL conical tube in a tube rack.", "[Concentration Procedure] Vigorously shake the sample to re-suspend solids. Immediately move on to Step 3.", "[Concentration Procedure] Add 50 mL of wastewater to the conical tube inside the biosafety cabinet.", "[Concentration Procedure] Transfe...
67,414
https://www.facebook.com/PureKanaKetoGummiesShop/
3
dx.doi.org/10.17504/protocols.io.e6nvwk3pwvmk/v1
https://www.protocols.io/view/https-www-facebook-com-purekanaketogummiesshop-cd3ws8pe
antoniocsmith
TITLE: https://www.facebook.com/PureKanaKetoGummiesShop/ AUTHORS: antoniocsmith [DESCRIPTION] These gummies are herbal weight reduction supplements that is to help you trim down your weight naturally without any unfavourable effects on your health. [STEPS]
[]
57,781
Amplifying the target genomic region by PCR
4
dx.doi.org/10.17504/protocols.io.b4nvqve6
https://www.protocols.io/view/amplifying-the-target-genomic-region-by-pcr-b4nvqve6
Hanqin Li, Yogendra Verma, Dirk Hockemeyer, Frank Soldner
TITLE: Amplifying the target genomic region by PCR AUTHORS: Hanqin Li, Yogendra Verma, Dirk Hockemeyer, Frank Soldner [DESCRIPTION] This protocol describes a procedure for amplifying targeted genomic regions using a PCR reaction [STEPS] 1. For a 150bp paired-end or single-read sequencing experiment, design primers ...
["For a 150bp paired-end or single-read sequencing experiment, design primers in a way that the region of interest, like CRISPR cutting site or specific mutations, locates within 130bp from at least one of the primers", "Add NGS adapters to 5’ end of primers following the instructions of your local NGS facility", "20 µ...
98,168
Mo&#39;orea Surf Tourism Research Protocol
0
dx.doi.org/10.17504/protocols.io.kqdg3258qv25/v1
https://www.protocols.io/view/mo-39-orea-surf-tourism-research-protocol-db4y2qxw
Flynn Ryan Dartland
TITLE: Mo&#39;orea Surf Tourism Research Protocol AUTHORS: Flynn Ryan Dartland [DESCRIPTION] This protocol describes a method of understanding the attitudes of Mo'orea locals towards the development of surf tourism on the island of Mo'orea, French Polynesia. The research involves interviewing local surfers, residents,...
["[Interviewing Surfers] The first step is to identify surf breaks on Mo’orea. To locate surf breaks, you can use a map to identify reef passes where waves break, the Surfline app, the internet, and/or ask locals in the area about where surf breaks are located.", "[Interviewing Surfers] Paddle, kayak, or boat out to a ...
39,102
Two-Bottle Choice Protocol
1
null
https://www.protocols.io/view/two-bottle-choice-protocol-bie6kbhe
Marcella Hughes, Olivier George
TITLE: Two-Bottle Choice Protocol AUTHORS: Marcella Hughes, Olivier George [STEPS] ?. [24-Hour Two Bottle Choice Procedure] Record weekly body weights of animals for calculation of solution intake as mg/kg ?. [24-Hour Two Bottle Choice Procedure] Mix and fill required solutions in bottles ?. [24-Hour Two Bottle Choice...
["[24-Hour Two Bottle Choice Procedure]\nRecord weekly body weights of animals for calculation of solution intake as mg/kg", "[24-Hour Two Bottle Choice Procedure]\nMix and fill required solutions in bottles", "[24-Hour Two Bottle Choice Procedure]\nRecord weight of all bottles prior to drinking session", "[24-Hour Two...
null
null
null
dx.doi.org/10.17504/protocols.io.hmgb43w
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>We recently developed a continuous flow system to transform microorganisms in high throughput in a microfluidic device (Garcia et al., 2017). This system employs microfluidic channels that contain a bilateral constriction between the inlet and outlet electrode connections (<e...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.j7hcrj6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><strong>INTRODUCTION: </strong>Blindness is and, apparently always has been, a problem in Egypt. Corneal blindness is a major public health problem worldwide according to the World Health Organization (WHO), which estimated that in every year, about 1.5–2.0 million new cases ...
[]
77,215
DeRisi Lab RNA Library Prep 96-well Protocol on Echo 550
1
dx.doi.org/10.17504/protocols.io.n2bvj8jrngk5/v1
https://www.protocols.io/view/derisi-lab-rna-library-prep-96-well-protocol-on-ec-cpm7vk9n
Madeline Y Mayday, Miriam R Simon, Lillian M Khan, Eric D. Chow, Matt S Zinter, Joseph Derisi
TITLE: DeRisi Lab RNA Library Prep 96-well Protocol on Echo 550 AUTHORS: Madeline Y Mayday, Miriam R Simon, Lillian M Khan, Eric D. Chow, Matt S Zinter, Joseph Derisi [DESCRIPTION] Preparation of high-quality sequencing libraries is a costly and time-consuming component of metagenomic next generation sequencing (mNGS...
["Spin plate of sample RNA in vacuum evaporator at the appropriate temperature and time settings to dry completely (approximately 37-38ºC for 1 hour, depending on number of samples and machine used).", "[GeneVac] Warming up GeneVac: Turn to \"aqueous\" setting and set temperature to ~40-42ºC and let run for ~30min befo...
91,462
jres_metab.nan
5
dx.doi.org/10.17504/protocols.io.e6nvwd27wlmk/v1
https://www.protocols.io/view/jres-metab-nan-c5jey4je
NAN KB, John Glushka, Mario Uchimiya, Saraa Al Jawad, Christopher Esselman, Leandro I Ponce, Laura Morris, Arthur Edison
TITLE: jres_metab.nan AUTHORS: NAN KB, John Glushka, Mario Uchimiya, Saraa Al Jawad, Christopher Esselman, Leandro I Ponce, Laura Morris, Arthur Edison [DESCRIPTION] This is a protocol for running the Bruker pulse program "jresgpprqf". [BEFORE_START] This protocol assumes your sample is loaded, locked, tuned, and shi...
["[Create a new dataset]", "[Create noesypr1d experiment file] When Lock, Tune, Shim are complete proceed to the specific protocol for your desired pulseprogram(s).", "[Create a new dataset] On the menu bar on TopSpin, click on\nStart → Create Dataset", "[Create a new dataset] Enter\nNAME: Name of a set of datasets (e....
28,349
Viral metagenomics using SMART-9n amplification and nanopore sequencing
null
dx.doi.org/10.17504/protocols.io.7w5hpg6
null
Ingra Claro Morales, Josh Quick
TITLE: Viral metagenomics using SMART-9n amplification and nanopore sequencing AUTHORS: Ingra Claro Morales, Josh Quick [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is a SMART-Seq (Switch Mechanism at the 5′ End of RNA Templates) protocol developed to use random 9n priming and be compatible ...
["[Centrifugal filtration]\nTransfer up to sample directly onto a Ultrafree-MC Centrifugal Filter column\n500 µl", "[Centrifugal filtration]\nSpin at for\nCentrifuge: 5000 34", "[Centrifugal filtration]\nRecover filtrate into Eppendorf tube\n1.5 ml", "[Centrifugal filtration]\nRemove basket and discard", "[Centrifuga...
103,467
Human Neuronal Nucleus Isolation for Single-Nucleus Transcriptomic Profiling (10x Genomics)
0
dx.doi.org/10.17504/protocols.io.4r3l2q3p4l1y/v1
https://www.protocols.io/view/human-neuronal-nucleus-isolation-for-single-nucleu-dhaj32cn
Satoshi Ishishita, Allan-Hermann Pool
TITLE: Human Neuronal Nucleus Isolation for Single-Nucleus Transcriptomic Profiling (10x Genomics) AUTHORS: Satoshi Ishishita, Allan-Hermann Pool [DESCRIPTION] Protocol generating suspensions of neuronal nuclei (NeuN+) from human central nervous system tissue for single-nucleus transcriptomics. [STEPS] SECTION: Equip...
["[Equipment and Reagents] Equipment\nKimble Dounce Kontes tissue-grinder set (DWK 885300-0000)\n50 ml Oakridge tubes (#0556214D)\n15 mL Falcon tubes (Fisher #352097)\n50 mL Falcon tubes (Fisher #352070)\n1.5mL LoBind Eppendorf Tubes\n70-micron Corning Cell Strainer (#431751)\nFire polished glass Pasteur pipettes (VWR ...