... our 96-format pipeline, we can routinely achieve 5-30% tag knock-in efficiency across different stem ...other suppliers with a typical turnaround time of 5-7 days. The crRNA and tracrRNA are annealed on a PCR...into stem cells. Cells are allowed to recover for 2-5 days and then analyzed for tagging efficiency using... in 96-well format using an Amaxa Shuttle device; 5 days after the transfection, the entire plate was ...commercial suppliers of CRISPR reagents (turnaround time 5-7 days); because the tracrRNA is the same in all csRNP...suspension, transfect and let cells recover for 2-5 days.Note: We use the Nucleofection method for delivery...J Biotechnol 241: 136-146. PubMed PMID: 27845164. 5. Clarke, Ryan, et al. "Enhanced bacterial immunity...

...MagIC-cryo-EM to determine several structures (at < 5 Å resolution) of the linker histone H1.8 in Xenopus...edited sequence anneal with each other, while the 5’ flaps containing the original DNA sequence anneal...anneal with each other. Excision of the annealed 5’ flaps yields an edited sequence with the attB/P landing...with twin prime editing. Nature Biotechnology, 40(5), 731–740. doi: https://doi.org/10.1038/s41587-021...8R] antibody shared by James Trimmer’s lab (Figure 5) as well as antibodies for identifying other cell ...anti-GFAP [N206A/8R] antibody here!   Figure 5: Free-floating rat brain sections stained with Anti-GFAP...transcriptional silencing (Nuñez et al., 2021). Now, in a collaboration with the Vallabh/Minikel Lab, the Weissman Lab...

...iPSCs from PBMCs. Most laboratories use Sendai virus to reprogram PBMCs efficiently [5]. However, the Sendai...establishment of virus-free hiPSCs usually require about 5-10 passages. Furthermore, working with Sendai virus...2003. 302(5644): p. 415-9. PubMed PMID: 14564000. 5. Seki, T., S. Yuasa, and K. Fukuda, Generation of ...pluripotent stem cells. Korean J Intern Med, 2014. 29(5): p. 547-57. PubMed PMID: 25228828. 7. Okita, K., ...p53-p21 pathway. Nature, 2009. 460(7259): p. 1132-5. PubMed  PMID: 19668191. PubMed Central PMCID: PMC2917235...source of somatic cells. This translates to only about 16 hiPSC colonies per 100 ml of starting blood sample...

... Hive Beautifully filmed video about colony collapse disorder.     5. Plasmid Rap Plasmid rap video...Science with Tom. In this podcast, we learn more about Tom and pick his brain for advice on how to dive... Rap of Henrietta Lacks Students in Oakland rap about Henrietta Lacks - the African American woman whose...science standards and communicate science. Learn more about Tom on his youtube channel and at sciencewithtom.com...

... for 5 years and just recently stepped down. I have gained invaluable experience and even after 5 years...Addgene executive director Joanne Kamens has written a 5-part blog on the topic:  http://blog.addgene.org/management-for-scientists-managing-vs-leading...in as Chair of the Committee which I then led for 5 years.  Different leadership styles Being an effective...the work I agreed upon (just as I would in my laboratory work). Committee members have to work professionally...to work with many different types of people both above and below you. During my time at the NPA, there ...further opportunities opened up to me. As I spoke about my work with the NPA, I was ultimately invited to...society in my scientific field - ARVO (as mentioned above). I became an official member of the Committee and...

...Exo is a 5’→3’ dsDNA-dependent exonuclease.  Exo will degrade linear dsDNA starting from the 5’ end and...alternative approach is to flank the desired change with 4-5 silent changes in the wobble codons, i.e. make changes...changes to the third base pair of the adjacent 4-5 codons that alter the nucleotide sequence but not the... of the translated protein.  These changes can be 5’ or 3’ of the desired change.   Table 1: Overview ...PMID: 19180090. PubMed Central PMCID: PMC2790811. 5. Thomason, L. C., Sawitzke, J. A., Li, X., Costantino...currently a research scientist in the Department of Laboratory Medicine at the University of Washington. She... Additional Resources on the Addgene Blog Read about Homology Directed Repair Use Knockout/Knockin Plasmids...

... expressing Rep2 and Cap2 Melina Fan 104964 pAAV2/5 AAV5 AAV packaging plasmid, expressing Rep2 and Cap5...production. Read our AAV Guide for more information about: AAV components Common uses of AAV Viral integration...out our Viral Vector Packaging Service and learn about our options for hundreds of in-stock preps that ...

...proposal successful. Typical proposals consist of 3 to 5 aims, with secondary milestones to meet along the ...statements on the next round of applications. Step 5: Keep writing! Each written grant application builds...first. Additionally, reach out to peers and collaborators who have succeeded at grant writing to see what...

...retrograde transport has been found for AAV1, -2, -5, -7, -8 (Taymans et al., 2007), AAV1 (Hollis et al...Anterograde transport has been observed for AAV1, -5, -8 (McFarland et al., 2009), and bidirectional transport...tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One 8:e76310....Improved Vectors for Gene Delivery. Curr Gene Ther 5(3), 299–310. PubMed PMID: 15975007. PubMed Central...Concepts in the pathogenesis of rabies. Future Virol 3(5), 481–490. PubMed PMID: 19578477. PubMed Central PMCID...transduction efficiency of recombinant AAV serotypes 1, 2, 5 and 8 in the rat nigrostriatal system. J Neurochem... of adeno-associated viral vector serotypes 1, 2, 5, 7 and 8 in mouse brain. Hum Gene Ther 18, 195–206...

... cleaves in a staggered fashion, creating a 5 nucleotide 5’ overhang 18-23 bases away from the PAM. Cpf1...some of Cas9’s shortcomings. Cpf1’s preferred PAM is 5’-TTN, differing from that of Cas9 (3’-NGG) in both...evolutionary classification of CRISPR-Cas systems. 5. Nat Rev Microbiol. 2015 Sep 28. doi: 10.1038/nrmicro3569...a cell culture model. What we still don't know about Cpf1 In their original paper characterizing Cpf1...Addgene Blog Catch Up On Your Cpf1 Background Read About CRISPR Competitor NgAgo Learn How You Can Use piggyBac...

...in Malawi 4:36-5:46 : How Kingdom ended up working in the life sciences and agriculture 5:47-9:35 : Kingdom's...more about Dr. Kwapata’s current work and thoughts on the need for more scientific collaboration across... connecting us with him. Learn more about our recent collaboration with Seeding Labs here. Special thanks...received an award from nonprofit and Addgene collaborator, Seeding Labs, for lab equipment that will help...Addgene Podcast.     Additional Resources Learn about Seeding Lab's Work with Lilongwe Univeristy of Agriculture...How You Can Contribute to the Addgene Blog Learn about Addgene Plasmid Donations for Rare Disease Research...

..., and a room of 5-30 faculty members. The session lasts up to an hour. You will get 5-10 minutes to present... should be mechanistic, doable by 1-3 people in 1-5 years, and contribute to resolving the overarching...solve the problem. What will you have accomplished in 5-10 years? You need to be able to describe what success...a single grant. Either way, think realistically about the time and resources required to feasibly accomplish...interrupted frequently by committee members. As mentioned above, plan to give your 10 minute overview and address... list of common questions, check out this eBook about applying for faculty positions. You need to practice...program. You should be proud of it and be excited about getting the opportunity to realize your vision. ...

...was provided by Addgene on 5/26/20 and contains requests from 5/22/14 to 5/19/20). Each paper has between...break, and COVID-19 on plasmid requests Figure 5: Inferred effects. Values are the inferred percentage...in his laboratory. Learn more here. Additional resources on the Addgene blog Learn more about plasmid...

...PHP.B. Gene Therapy, 25(5), 392–400. https://doi.org/10.1038/s41434-018-0028-5 de Backer, M. W. A., Brans...We found that, in this case, AAV1 (23%) and AAV6 (<5%, data not shown) were not good options while AAV8...Initiative, F. B. (2019). Setting your data free. Nature, 5–7. O’Connor, D. M., Lutomski, C., Jarrold, M. F., ...asked why they don’t share, many express concern about sharing negative data they cannot explain, perhaps...

...alternating between 5 min at -20°C (or -80°C) followed by 5 min at 37°C (each repeated 5 times) works well...without the need for genome sequencing or annotation (5). In the event that genome sequencing can be done,...Sacher, and Szymanski in January 2016 in Viruses (5). Please refer to the paper for more details on the...our planet, but we still know collectively little about how they infect and influence the evolution of their...

... Trypanosoma brucei Cross 34564 pMSCVpuro-Cre Cre 5'LTR Retroviral Ge 34565 pMSCVhygro-Cre Cre and HygroR...HygroR coexpression Retroviral Ge 34568 WZLneo-Cre Cre 5'LTR Retroviral Ge 35619 pTAT-CRE Cre fused to HIV-...1027-32. doi: 10.1073/pnas.0610155104. Epub 2007 Jan 5. PubMed . Sauer B, Henderson N. 1988. Site-Specific...Transgenes. Proc Natl Acad Sci. Jul 13;101(28):10380-5. doi:10.1073/pnas.0403954101. Epub 2004 Jul 6. PubMed...Learn about the Cre-Lox system and how it can be used for genetic rearrangements and recombination. Use...

...09 - 5:58 : Specific Examples where Niroshi and Ben helped build their grad student communities 5:59 -...How did you hear about post grad opportunities and when did you start thinking about them? 18:30 - 19:... top if you leverage your community, think hard about picking a good mentor, and begin considering career...19:54 : How to think about picking a post-doc project 19:55 - 22:40 : Improvements that can be made to ...

...take to fuel more than 30,000 neuroscientists over a 5 day conference? I feel like here at the annual Society...of coffee would be consumed throughout the entire 5 day event. Though, to be fair, I think this is a conservative...learn about all the amazing research being done with plasmids. I heard an impressive talk about the use...pulls off such an inspiring, thought-provoking, collaborative event – all while keeping the party going! At...scientists we've spoken with are excited and passionate about making such tools open-access. Though it is time...

...derived from the P and V proteins of Simian virus 5 (SV5) (Randall et al., 1987). The epitope (GKPIPNPLLGLDST...characterization of monoclonal antibodies to simian virus 5 and their use in revealing antigenic differences between...assemble and catalyze the luminescence reaction (Figure 5). The researchers used the reporter to visualize biologically...cells in several different contexts.   Figure 5: Left: Mechanism of the PILATeS tension sensor. The...more feasible, the lab of Alessio Ciulli and collaborators developed CRBNmidi (Kroupova et al., 2024). ...stepwise, achieving mutation efficiencies of up to about 9% of colonies (102 to 104 times higher than traditional...Bentley Time to get flexible! Alex Dunn’s lab and collaborators have deposited a new mechanosensor called PILATeS...

...rmmAkm6PQd — Elizabeth Kenkel (@ElizabethKenkel) December 5, 2018 How social media has helped me in science...have tips for using social media as a scientist. 1. "5 Ways Scientists SHOULD Be Using Twitter" podcast from...include in each post I write for the Addgene blog. 5. "Social Media in Your Working Day" video from Science... advantage. It wasn’t until I wrote a blog post about a research paper that explored how scientists connect...saw a request for an Addgene guest blogger post about colony PCR, I signed up immediately. I had used ...science communication skills, while also learning about new scientific developments and helping scientists... found this paper. I wrote an Addgene blog post about it and shared the paper with my lab since one of...