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TypeBlog Post...tested was constantly tracked with barcodes and spreadsheets. It was great to be in the lab doing experiments...was to fulfill contracts, my work sometimes felt rushed and the focus always seemed more on getting results...
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Antibody Guide
TypeGuide...the number of signaling molecules that can be distinguished in your application. The indirect detection ...Cross-link the samples with formaldehyde or UV light. Shear the samples using sonication to break DNA up into...protein. Micrococcal nuclease digestion is used to shear the DNA in these assays. This method is suitable...specific to your antibody and assay may already be published. If it is not, you’ll need to validate the antibody... -
Pipetting Protocol
TypeProtocol...pipette, and the tip ejector ring at the bottom which pushes the pipette tip off of the pipette. This side view...rate into the pipette tip. Once the liquid has finished drawing into the pipette tip, lift the pipette... -
Transfection for Recombinant Antibodies
TypeProtocol... system, 500 mL, VWR 430770 Trypan Blue, Thermo Fisher T10282 Valproic acid sodium salt, Sigma Aldrich... Grade, VWR 75800-188 10% Pluronic F-18, Thermo Fisher 24040032 Glutagro, Corning 25-015-CI BalanCD HEK293... -
Antibodies 101: Antibody Engineering and Directed Evolution
TypeBlog Post...This post was written by Alyssa Shepard and Angelo Nicolaci, a PhD student at Moffitt Cancer Center. ... Alyssa Shepard... -
An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency
TypeBlog Post...Originally published Jan 27, 2015 and last updated Dec 17, 2020. This post was contributed by Jordan ...experimentation. In 2014, several manuscripts published in Genetics detailed approaches in which selection... -
9 tips for a successful postdoctoral experience
TypeBlog Post...Multiple first or corresponding author publications (published, not in preparation) A history of successful funding... meet future collaborators and get access to unpublished information and reagents. Part of the reason ... -
Sequencing Options for CRISPR Genotyping
TypeBlog Post...23934178. PubMed Central PMCID: PMC3782611. 8. Tsai, Shengdar Q., et al. "GUIDE-seq enables genome-wide profiling...23503052. PubMed Central PMCID: PMC3651036. 15. Tsai, Shengdar Q., and J. Keith Joung. "Defining and improving... -
Affinity Purification of Recombinant Antibodies with Protein A or Protein G
TypeProtocol...luer-lock), VWR 431229 IgG Elution Buffer, Thermo Fisher 21028 Protein G GraviTrap columns, Cytiva 28-9852...Spin Desalting Columns, 7 kDa MWCO, 10 mL, Thermo Fisher 89893 Amicon Ultra-15 30 kDa MWCO column, Millipore... -
Isolating a Monoclonal Cell Population by Limiting Dilution
TypeProtocol...of adherent cells) Trypsin EDTA (TrypLE, Thermo Fisher 12605-010) 6-well tissue culture treated dish, ...trypsinize the cells and expand them to larger culture dishes, being sure not to cross contaminate the independent... -
Kit Free RNA Extraction
TypeProtocol...opens in a new window) TRIzol® User Guide from ThermoFisher Scientific . Figure 1: A diagram of the different... Caution Make sure to read the SDS (Safety Data Sheet) for safety warnings and hazards for these reagents... -
CRISPR 101: Multiplex Expression of gRNAs
TypeBlog Post...Originally published Jan 28, 2016 and last updated Sep 10, 2020 by Jennifer Tsang. CRISPR makes it easy...plasmids from this paper at Addgene. 5. Tsai, Shengdar Q., et al. “Dimeric CRISPR RNA-guided FokI nucleases... -
Antibody Validation for Flow Cytometry
TypeBlog Post...manufacturer and/or in the published literature. Note that the antibody being published doesn’t always indicate... -
AAV Production in HEK293 Cells
TypeProtocol...) Opti-MEM, Thermo Fisher 31985-070 0.05% Trypsin/EDTA (e.g. TrypLE, Thermo Fisher, 12605010) 1X PBS pH... -
Lentivirus ddPCR Titration
TypeProtocol...Reagents GeneJet Genomic DNA Purification Kit, Thermo Fisher, K0721 6-well tissue culture treated dish, VWR,...of adherent cells) Trypsin EDTA (TrypLE, Thermo Fisher, 12605010) Ethanol, VWR, EX0276-1 Benzonase 250... -
Using AAV for Neuronal Tracing
TypeBlog Post...advantage of AAV vectors is that there are well-established protocols for their production and purification...PubMed Central PMCID: PMC4324159. Castle, M.J., Gershenson, Z.T., Giles, A.R., Holzbaur, E.L., and Wolfe... -
Adenovirus Guide
TypeGuide... you're new to viral vectors or simply need a refresher, we've included a glossary at the end! Browse ...remain the most popular choice due to their well-established methods, relative simplicity of production, and... -
Adeno-associated virus (AAV) Guide
TypeGuide... you're new to viral vectors or simply need a refresher, we've included a glossary at the end! Browse ..., and VP3, which together form the outer capsid shell that protects the viral genome. Helper plasmid E4... -
New Antibodies in the Lab Animation!
TypeBlog Post...cytometry. It's a great introduction to — or quick refresher on — these common antibody applications. Thanks... -
Introducing...Addgene Quick Tips!
TypeBlog Post...is below. And you’ll be able to find all published videos in our Quick Tips playlist: Want your...