...expected to grow with the ever-expanding genetic toolbox generated by the continuous effort of numerous scientists...). GFP is the most popular, most widely used genetically encoded fluorescent probe. Several factors contribute...probe, it is the first choice when selecting a genetically encoded fluorescent tag. There are, however, ...1998). Another mutation (A206K) is necessary to generate the strictly monomeric EGFP variant, mEGFP (Zacharias..., followed by an intramolecular reaction that generates the chromophore (Tsien, 1998). Importantly, this...Thorn, 2017). These fluorescent analogues are generally added to the bathing solution and require uptake...adding GFP may alter diffusion rates. Smaller genetically encoded probes are based on cofactor binding ...

...If you’ve ordered a pooled library from Addgene, you may have been a little surprised at how this powerful...deposited many types of pooled libraries here at Addgene, including cDNA, barcoding, screening, and gRNA...gRNA libraries. Most of the pooled libraries at Addgene are lentiviral pooled libraries, though other types...types exist.  When you buy a pooled library from Addgene, you can either order a DNA pooled library, which...single tube, or a pre-made viral prep library. Addgene’s pooled libraries are typically DNA preps, but ... A) Amplification, validation, and lentiviral generation of a pooled library. An aliquot of the pooled...experiments. Pooled library lentiviral preps are generally aliquoted to reduce the number of freeze-thaw ...

... institution’s TTO or legal office first. Generally, Addgene is also waiting to receive that legal agreement...materials from Addgene can qualify. We also reward improvement against one’s own record Mention Addgene Master...question? Refer them to Addgene’s Tech Transfer Team: techtransfer@addgene.org Congratulations to the...between a TTO and Addgene, whose terms apply to all future deposits of materials with Addgene Master Reception...between a TTO and Addgene, whose terms apply to all future materials requests from Addgene MTA: material ...commercialization Additional resources on Addgene.org Addgene Help Center: Tech Transfer Technology Transfer...arrival of the newest plasmid you requested from Addgene...the plasmid that will help you run that last ...

... Resources on Addgene.org Learn how Addgene engages with the community on our Addgene gives back page...Initiatives for the Personal Genetics Education Project (pgEd). Advances in genetic technologies and other ...broadens the conversation about genetics pgEd, based at the Department of Genetics of Harvard Medical School...existing inequalities and injustices. The Personal Genetics Education Project (pgEd), where I currently work...work, strives to broaden the conversations about genetics and facilitate dialogue between scientists and...the personal, social and ethical implications of genetics. In these conversations, pgEd strives to be inclusive...and empowered to make their own decisions about genetics, both as individual consumers of technologies ...

...Resources at Addgene.org Find pooled libraries at Addgene Find basic lab protocols at Addgene ...As a tech at Addgene, part of my job, in addition to preparing DNA and generating/replenishing kits, ...information from the workshop. Future workshops for Addgene Addgene strives to be a great resource for the research...Technician at Addgene who is interested in medicine and music.       Additional Resources on the Addgene Blog ... use what we learn to improve processes in the Addgene lab. In this post, I’ll tell you a little bit about...excited to go there because both LabCentral and Addgene aim to make scientific advancement easier. NEBNext...extract libraries from previously fragmented DNA.The general process for library preparation using this kit ...

...your desired edit are all you need to generate a knock-in.   General considerations for designing a repair.... A Mouse Geneticist's practical guide to CRISPR applications. Singh P, et al. Genetics 114.169771; ...accurate mechanism for DSB repair. The process is generally error-free provided the DNA template used for ... DNA to the target sequence to be modified. In general, the insertion sites of the modification should...bases can be difficult, thus dsDNA plasmids are generally preferred for large insertions such as fluorescent...efficiency with circular plasmid templates is generally low; to increase the frequency of edits, researchers...targeting region from the vector. Alternatively, PCR-generated or commercially synthesized linear dsDNA templates...

...Libraries Resources on Addgene.org Addgene's Guide to Plasmid Pooled Libraries  Addgene's Pooled Library Collection...chemical transformations are generally cheaper and easier. One library that Addgene distributes with a chemical...contact us at deposit@addgene.org. Happy amplifying! More resources on the Addgene blog Genome-Wide Screening...If you’re depositing a pooled library with Addgene, you may be surprised to learn that we ask for an ...recombination and loss of plasmid diversity, so Addgene strives to distribute aliquots of the original ...requesting scientist receives a pooled library from Addgene, they’ll need to amplify the sample for further...developed for each individual pooled library; generalized amplification protocols tend not to work as well...

... CRISPR, Addgene gave to me: a primer on prime editing.  On the second day of CRISPR, Addgene gave to ...On the fourth day of CRISPR, Addgene gave to me: ways to use optogenetics to control CRISPR editing.  ... day of CRISPR, Addgene gave to me: updates on base editing from the first generation of base editors ...the eighth generation (and dual base editors, too!).  On the seventh day of CRISPR, Addgene gave to me... of CRISPR, Addgene gave to me: anti-CRISPR proteins!  On the ninth day of CRISPR, Addgene gave to me:...🎶 On the 12 days of CRISPR, Addgene gave to me… 🎶 A few years ago we used the #12DaysOfCRIPSR hashtag...detect nucleic acids.  On the third day of CRISPR, Addgene gave to me: tips and tricks for designing gRNAs...

...no weak links in the chain.  Addgene: What about the Streptococcus pyogenes protein domain made it ideal...cover it. Addgene: OK, thanks for your time, and thanks for depositing your plasmids with Addgene. Howarth...lab at the University of Oxford is dedicated to generating new tools to manipulate biology based on molecular...based on a protein isolated from Streptococcus pyogenes that locks itself together, to produce irreversible...another important step forward by dissecting that S. pyogenes protein into three parts. Their efforts yielded... from the “S” in Streptococcus and the “py” in pyogenes), capable of locking two peptide tags together... and its potential uses. Developing a new tool Addgene: Where did you get the idea to develop SpyLigase...

...reference, Addgene commonly prepares NGS samples using both pre- and post-amplification DNA and generates PCR...CRISPR Libraries Resources on Addgene.org Browse Pooled Libraries at Addgene ...In addition to single plasmids, Addgene also distributes pooled plasmid libraries containing hundreds...million plasmids. These libraries are some of Addgene’s most exciting and versatile offerings! We recently...plasmids are amplified and none are lost. Each Addgene library has its own depositor transformation and...components of the library are represented. Typically, Addgene uses electrocompetent Stbl4 cells when performing...transformation efficiency. For a typical library, Addgene usually does four to eight electroporation reactions...