...Fill out our guest blogger form to get started!   Thanks for all your hard work!   Fluorophores When is...Casilio is no longer available through Addgene.  Thanks again to all of our fantastic depositors and guest...

...thermostable ligases and reaction buffers. Many thanks to our guest blogger, Greg Lohman! Greg Lohman ...

...contributed by the Developmental Studies Hybridoma Bank. Antibodies are the most widely used class of protein-binding...immunoprecipitation and more. The Developmental Studies Hybridoma Bank (DSHB), directed by David R. Soll, is a repository...Development (NICHD) in 1986, to enable basic research by banking and distributing monoclonal antibodies at cost....quality-control and technical support to the community. Thanks to Our Guest Blogger!   David Soll has been the... Director of the Developmental Studies Hybridoma Bank (DSHB) since 1996.       More Blog Posts about Scientific...

...fused to a fluorescent protein. “Frankenbody,” like the infamous frankenstein, was constructed by grafting...called this probe the HA frankenbody (Zhao et al., 2019).  To develop HA frankenbody, the Stasevich lab tested...can you do with the HA frankenbody? Another large advantage of the HA frankenbody is that it remains bound...provides HA frankenbody the ability to bind nascent HA tagged proteins.  Lastly, the HA frankenbody can be ...properly once inside cells.  Developing the HA frankenbody To combat the issues with these antibody based...lab observed colocalization of GFP tagged HA frankenbody with an mCherry, 4xHA tagged histone protein ... in U2OS cells. They then showed that the HA frankenbody probe could also bind a 1xHA tagged histone protein...

...-HA frankenbodies include mEGFP, mCherry, and HaloTag. Figure 1: (a) Anti-FLAG frankenbody (FB)...Here's what you'll find in this post: Anti-FLAG Frankenbodies for modular imaging of cellular targets Optimized...mammalian genome regulation and editing Anti-FLAG Frankenbodies for modular imaging of cellular targets by: ...: Mike Lacy Adding to their set of anti-HA frankenbodies (Zhao et al. 2019), Tim Stasevich and Hiroshi... have recently published a set of anti-FLAG frankenbodies, a flexible, modular tool for labeling. Here...variety of applications. This set of new anti-FLAG frankebodies includes mEGFP, mRuby2, iRFP713, SNAP-tag, and...Liu et al. 2021. Anti-FLAG and anti-HA frankenbodies have a variety of applications, including live-cell...

...the phage integrates into the bacterial genome flanked by two new recombination sites (attL-left- and ...BP Reaction takes place between the attB sites flanking the insert and the attP sites of the donor vector...the entry clone containing the DNA of interest flanked by attL sites. As a byproduct of the reaction, ... LR reactions. The BP reaction creates an attL-flanked entry clone. The LR reaction creates an expression..., an expression clone with the DNA of interest flanked by attB sites is generated. As in the BP reaction... generate our desired entry clone - human KRAS flanked by attL sites.  Method A: recombination of an attB-PCR...create an entry clone: recombination of an attB-flanked PCR product with an attP-containing donor vector...

...summary per batch. Ranking guides for HDR in-depth In other tools, guides are ranked by specificity of ...genome by eye. We came up with a ranking formula that does just this: Rank = (compressed CFD score) * (gaussian...employs biological best practices. For example, it ranks guides by CFD score, and it mutates HDR donor templates...between 0 and 1) so that they can be used in the rank equation above. Cut-to-insert distance was fitted...in the graph below) so that it can be used in the rank equation above. Generally, the greater this distance...chanzuckerberg.com. We're eager to hear your thoughts. Many thanks to our guest blogger, Greg Dingle! Greg Dingle ...

...innovation between legal and scientific fields.” Thanks to our legal team, and great customer service, ...and our visibility is increasing internationally, thanks to evolving outreach tactics, increasing business... have a UI/UX team. Last time I checked we were ranked as the 21,806th most popular website in the world...Directors is mature and poised to support our growth. Thanks to Joanna Brownstein (our outgoing Chair) and Dave... incorporate additional Board best practices. I thank them wholeheartedly for their service and support...excellent intentions, or incredible drive to succeed.  Thanks to all the Addgenies and congratulations to current...current and alumni Addgenies on our success. Thanks to our amazing team, we also don’t have to worry about...

...outside of their recognition sequence, creating flanking overhangs. These overhangs can be customized to...vectors each contain Type IIS restriction sites flanking the promoter-gRNA construct, but with different...with the appropriate Type IIS enzyme, the unique flanking overhang sequences can link together to allow ...plasmids contain Type IIS restriction sites that flank the promoter-gRNA constructs. When these plasmids...promoters, the gRNAs are transcribed together and are flanked by specific sites that allow them to be cleaved...nuclease system to cleave the gRNAs. gRNAs are flanked by glycine tRNAs to create polycistronic glycine...CRISPathBrick plasmid contains a nontargeting spacer flanked by two CRISPR repeats. The spacer can be digested...

...in brackets and add the upstream and downstream flanking sequences to their input string (Figure 2). This...provides users with different designs of pegRNAs, ranked by their PRIDICT/PRIDICT2.0 score. For the best...editing strategies) based on a DeepSpCas9 score ranking (Kim et al., 2019). PRIDICT saves time and resources... Nicolas Mathis received his PhD from Gerald Schwank's lab at the University of Zurich (Switzerland),...L., Solari, C., Balázs, Z., Krauthammer, M., & Schwank, G. (2023). Predicting prime editing efficiency...Böck, D., Steensel, B. van, Krauthammer, M., & Schwank, G. (2024). Machine learning prediction of prime...

...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...