...copies the sequence from a repair template with flanking sequence homology for error-free DSB repair. NHEJ...MMEJ uses regions with 5-25 bp of microhomology flanking a DSB to repair DNA. The DNA ends are chewed back...figure above, short (5-25 bp) regions of homology flanking a double-stranded break yield precise deletion... is likely to introduce additional nucleotides flanking the insertion. If the DNA ends anneal incorrectly...5’ and 3’ microhomology to the insertion locus flanking the GFP-Puro cassette. Three double stranded breaks...The PITCh plasmid contains a GFP-Puro cassette flanked by 5' and 3' microhomology and PITCh-gRNA binding...

...reversible. Whereas the excision of a piece of DNA flanked by loxP/FRT sites is favored over its reintroduction...vectors, the DNA sequence that should be inverted is flanked by two pairs of target sites that differ in their...with identical sequence leads to inversion of the flanked DNA sequence and the interior target sites. In ... leaving behind a vector with the inverted DNA flanked by non-compatible target sites (Schnütgen et al...genes in specific cells at specific times. Many thanks to our guest blogger, Katrin Michel! Katrin Michel.... PubMed Central PMCID: PMC41457. 8. Schnütgen, Frank et al. 2003. “A Directional Strategy for Monitoring...

...the blog. Thank you! More resources on the Addgene blog 10 Great Guest Posts We're Thankful For Lucky...It’s a very thankful time of year for those of us at Addgene! And we can’t think of any better way to...to start off our holiday season than by thanking the twelve guest bloggers and subject matter experts ...experts who have contributed to our blog in 2024. Thank you to…   Marty LaFleur, who shared information about...

...service is successfully launched, I would like to thank some of the people and organizations who helped ...milestone. Viral service supporters We would like to thank the companies who supported this project with funding...viral preps. Visit our Sponsors and Supporters Page Thank you to all of our generous, sharing depositors. ...encouraging us to explore new directions. I would like to thank them for their time, dedication, advice and encouragement...this well ... because it did. So finally, a big thank you to the Addgenies who went the extra mile to ...

...tool that responds strongly to magnetic fields (Frank Hayward et al., 2024). They deposited their plasmid...when a small magnet was passed over the sample (Frank Hayward et al., 2024). The fluorescence intensity...the magnetic field is on. Image reproduced from Frank Hayward et al. 2024 under CC-BY license. They...the magnetic field is on. Image reproduced from Frank Hayward et al. 2024 under CC-BY license.   ...468. https://doi.org/10.1016/j.bpj.2019.01.003.  Frank Hayward, R., Rai, A., Lazzari-Dean, J. R., Lefebvre...

... Addgenies use our social media and blog to say thank you to all the many scientists and researchers who...12,427 plasmids deposited with us. We cannot say thank you enough - but this week we’re certainly going...   Here’s our Depositor Week schedule:  Monday: Thank you post (this is it!)  Tuesday: Stats roundup on...Addgene swag.  And, of course, to all our depositors: thank you so much!!!  (A special shout-out to those who...

...antibodies (mAbs). mAbs have been around since 1975 thanks to the Nobel prize winning work of Milstein and...single molecule tracking in living cells. The HA frankenbody is an HA tag detection probe. It contains an ...to a fluorescent protein such as GFP. The HA frankenbody works just like an antibody-based probe, but ... resources on the Addgene blog: Read about frankenbodies, an anti-HA scFv-based probe for fluorescent... antibodies, the Developmental Studies Hybridoma Bank Learn more about the SunTag system which uses an...

...the interview with Tom McFadden!   1. Rosalind Franklin vs. Watson & Crick - Science History Rap Battle...telling following the unfortunate story of Rosalind Franklin and her incredibly important role in solving the...Fill in the Planck at sciencewithtom.com. Many thanks to Tom McFadden for taking the time to sit down...youtube channel and at sciencewithtom.com.   Special thanks to our sound engineer/producer, Eddy Page! Edward...

...without exclusivity. Chapter 11: The Stem Cell Bank (CReM Bank) at the Center for Regenerative Medicine and...did I start doing this in the first place?" Many thanks to our guest, Darrell Kotton. Dr. Kotton is the...Boston University and Boston Medical Center. Special thanks to our sound engineer/producer, Eddy Page! Edward...

... 17] should cover it. Addgene: OK, thanks for your time, and thanks for depositing your plasmids with ...Howarth: Just the usual blood, sweat and tears. (Thanks to our healthy blood donor for keeping going while...be of use to others to Addgene. I am a big fan; thanks for the T-shirt! References: SpyLigase peptide-peptide...

...package. In a transfer plasmid, the transgene is flanked by two ITRs, and the ITRs are retained with the...Figure 1: Recombinant AAV vector with ITRs that flank the gene cassette, which contains a promoter, the...ITRs are symmetrical 145-nucleotide sequences that flank the ends of the single stranded DNA genome of AAV...the native virus. In an AAV vector plasmid, they flank the gene of interest. A single ITR consists of palindromic...

...results: On this page you can download an annotated GenBank File or a SnapGene file for the sequences listed.... If the depositor provided a custom annotated GenBank file, this can be found on the main plasmid page...relevant), and types. Types listed are standard GenBank feature types. Clicking on a particular feature...your next experiment. We would like to extend our thanks to you (our community of scientists) for suggesting...

...reproducibility and shared similar desires to take action. Thanks to Teytelman’s vast network and ability to identify...thinking about ways to address irreproducibility and thanks to Teytelman’s connection with Joanne Kamens, Addgene... them to connect? Through networking of course! Thanks to individuals in the group who were well connected...collaborate with this incredibly dedicated team. Many thanks to Tyler Ford, Caroline LaManna, Joanne Kamens,...

...didn't have the space to give proper credit to. Thankfully, as Addgene's resident blog editor, I could take...can refold correctly after denaturing.  Designed ankyrin repeat proteins, or DARPins, are genetically engineered...derived from a class of binding proteins known as ankyrin repeat proteins. DARPins typically consist of 3...an excellent option for many applications, and frankly, if ain't broke, don't fix it!  That being said...

...Harvard Medical School. We want to give a special thanks to the early-adopter scientists, who trusted Addgene...to hear the stories others have to tell.  "Also thanks for being Addgene. You guys rock." — Andrew York...Calico Life Sciences. Most of all, we’re incredibly thankful for all the scientists who have joined our community...your work, your research, and your discoveries. Thank you. You guys rock. ...

...the design of flanking overhangs. Although there are theoretically 256 distinct flanking sequences, sequences...their recognition sequence, creating four base flanking overhangs. Since these overhangs are not part ...can engineer unique enzyme recognition sites that flank their DNA fragment in an inverse orientation. This...

...recombination events, and that the orientation of the flanking loxP sites directs the Cre recombinase to invert...while FLP binds FRT sites. When a DNA sequence is flanked by target sites (floxed) in opposing orientations...Figure 1, top). The entire DNA cassette would be flanked by two pairs of target sites- one wild-type pair...placement of the two pairs of target sites. By flanking the reporter with one loxP site and one lox511...

...vector contains a neomycin resistance gene (NeoR) flanked by homology arms. The negative selection marker...” the resistance gene through the insertion of flanking LoxP sequences, you can remove the gene through...targeting vector such that a set of three LoxP sites flank the resistance gene and the targeted exon in the...has simply been replaced with the same sequence flanked by LoxP sites while the resistance cassette has...

...two sets of sgRNAs; A, the top 3 ranked sgRNAs, and B, the next 3 ranked sgRNAs. Dolcetto was able to discriminate... lab selected sgRNAs in this optimal window and ranked them based on their optimized sgRNA rules and potential... and Dolcetto library, sgRNAs were selected and ranked based on the previously established sgRNA rules...

...he describes the now infamous tale of Rosalind Franklin and her essential-but-unlauded role in the identification...within this section. The GFP story has its own Franklin in Doug Prasher, who originally isolated the GFP...science produced by the quieter, more reserved Franklins and Prashers will find it increasingly difficult...your career.  Acknowledgements: I would like to thank fellow Addgenies Joanne Kamens, Marcy Patrick, Aliyah...