...mind that access and availability to resources (including the internet!) can vary depending on factors like...your best to understand what kind of resources - including your time - will be required to produce an outcome...naked eye or to make things more accessible by including, highlighting, or omitting details. Digital or...

...viral vectors in the lab. Advantages of lentivirus include a large genetic capacity and the ability to transduce...CRISPR plasmids are designed for lentiviral use, including Addgene’s top requested plasmid in 2015: lentiCRISPR... observed in small trials for other diseases, including the common hematopoietic disorders sickle cell...

... and management process or the whole process, including grant readiness, funding opportunity research,...grant training, mentoring and coaching services, including Smartegrants, to help those entering the Grants...LLC, a grant professional development firm that includes the social media community GrantChat. She has ...

...improvements? You could publish a paper that happens to include your improvements, or maybe even a methods paper...variety of useful molecular cloning protocols, including making LB agar plates, restriction digest of plasmid...been known to recommend protocols listed on OWW, including those for preparing your own competent cells. ...

...scientists employ GFP for a wide number of functions, including: tagging genes for elucidating their expression...detect a variety of intracellular conditions, including ion (such as Ca2+) concentrations and pH, using...selection: Expression constructs like plasmids often include GFP as a marker to help identify which cells have...

...If you choose this strategy, it’s important to include the proper protein expression elements (ribosome...entry clones available for many popular genes, including Hs.KRAS4a. Use our website to search for your ...Gateway Multisite plasmids available at Addgene, including the Frew Lab Multiple Lentiviral Expression Systems...

...the CRISPR workflow. For a multiplex sample, ICE includes visual representations of all detected edit types...indel.   For a complete guide to ICE analysis, including detailed explanation and examples, read the paper...stack Genome Engineering Solutions, which also includes our CRISPR knockout guide design tool, guide validation...

...rather late stages (i.e. postdoc level) and can include the pressure to publish in high-impact journals...for insights from my time in technical support including tips, advantages, and disadvantages of this overall...without being too commercial or “sales-y”. The work includes reviewing customer data sets, troubleshooting ...

...contain components of the SB transposon system including those from the original publication, Ivics et ... been studied in many contexts within the fly including oogenesis and cytokinesis. Why name this protein...that contains bagpipe from the Deplancke lab. Conclusion I hope you enjoyed this tour of Addgene’s new...

...proteins. Figure 4: Reporters in Gene On include a mScarlet reporter, luciferase reporter, and neomycin...translation of an array of different reporters including mScarlet-I, luciferase, antibiotic resistance ...can be used for an array of detectable proteins including fluorescent proteins, antibiotic resistance, and...

...many types of pooled libraries here at Addgene, including cDNA, barcoding, screening, and gRNA libraries...viruses are sensitive to freeze/thaw cycles, we include a 1.3 mL aliquot with mostlibrary requests specifically...aliquot to full use! Common conditions to optimize include incubation time, antibiotic conditions, and amount...

... hardest part of any revolutionary discovery, including CRISPR, is turning potential into impact. In molecular...measurable cell toxicity – using Cas7-11. This included tests done in cell lines known to be more sensitive... — the ability to now make any small changes, including base edits, insertions, and deletions. But CRISPR...

...spacers of the crRNA can be customized as well, including concatenations into crRNA arrays targeting multiple...by sacB counterselection.  The MUCICAT toolkit includes versions of plasmids with different induction ...concentration. Image from the Yang lab.   In conclusion, the MUCICAT system has the following advantages...

...N-glycosylase (UNG) enzymes. Various other proteins included in different CGBE tools allow the researcher to...again, this post has a lot of acronyms, so we've included a glossary at the end of this post! Figure...for their application from among many options.  Conclusion With these advances, every base can now be directly...

... bioinformatics pipelining In many use-cases, including ours at Addgene, the overall purpose of a data...workflow management system for scientific pipelines, including bioinformatics. We loved the parallels between...language.  The modern architecture of Nextflow included built-in support for containers, such as Docker...

...the origin and propagation of the line, and can include assays for authentication and contamination) and...International Cell Line Authentication Committee includes a database listing all cross contaminated or misidentified...working for a variety of pharmaceutical companies including GSK, AstraZeneca-MedImmune, and Infors & Applikon...

...expression strains to accommodate high protein levels including:  ompT: Strains harboring this mutation are deficient...Rhamnose-tunable T7 RNAP expression alleviates inclusion body formation. Expression vector cannot have ...insoluble globs of misfolded protein are known as inclusion bodies, and can be recovered and purified using...

...have created synthetic promoters, which typically include some combination of other promoter elements, and...promoters for eukaryotes and prokaryotes We have included two reference tables below listing some of the... simian vacuolating virus 40 Constitutive  May include an enhancer. PGK1 (human or mouse) General expression...

...qualitative and quantitative measurement of processes including RNA movement by tagging RNA sequences with an ...Kellenberger et al., 2013). Other variants of Spinach include Baby Spinach, a miniaturized version of Spinach...aptamer fluorophores, more are in development, including Carrot and Radish from the lab that developed ...

...choose a looser gate here. I am not worried about including irrelevant events in my analysis, as these will...Lastly, you learned which gates should always be included in your analysis, regardless of your experiment...Sharpe at Harvard Medical School. His projects include investigating negative regulators of T cells and...