... 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...

... 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...

...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...

...that adding an additional guanine nucleotide to the 5’ of 20-bp guide can increase targeting efficiency....that adding an additional guanine nucleotide to the 5’ of 21-bp guide can increase targeting efficiency....Addgene Zhang Lab CRISPR Resources Tips for and FAQs about the Zhang Lab CRISPRs Browse All Zhang Lab Plasmids...

...gram-negative bacterial endotoxin is ensured to be less than 5 endotoxin units per mL. The endotoxin assay is carried... before being distributed to customers. Details about our production protocols, titering methods, and ...either in-house by Addgene scientists or through collaboration with viral vector manufacturing facilities, ...

...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...

...process (see figure above), a fragment of interest is PCR amplified to add the specified 5’ and 3’ homology...products are used to generate a target gene with a 5’ or 3’ overhang. When the products are mixed and annealed...similarity. If fragments in a multicomponent assembly have 5’ or 3’ sequence homology to each other, they may be...polymerase to create longer, “chewed-back” overhangs of about 10-12 bases. Only one type of dNTP would be present... or iPCR, fragments (right branch of the figure above). If PCR does not include a final extension step...recombination. Mixed PCR, shown in the middle branch of the above figure, can also be used to create an insert. Two...

...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...

...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...

...exo-) DNA Polymerase I, Large (Klenow) Fragment (3'-5' exo-) 165512 pOpen-THEAQpolA Taq DNA Polymerase 165513...engineering and directed evolution. More information about the collection can be obtained at the Open Bioeconomy...

...Jude JG, Zuber J, Gerlich DW Cell. 2017 Aug 24;170(5):956-972.e23 Screen identifies bromodomain protein...Jude JG, Zuber J, Gerlich DW Cell. 2017 Aug 24;170(5):956-972.e23 Rapid and phosphoinositol-dependent binding...Swiderek K, Kuo A, Crabtree GR Cell. 1998 Nov 25. 95(5):625-36. An architectural role for a nuclear noncoding...Leah Bury of Microscopic Art. I started thinking about this problem when I recently attended the 2018 Chromosome...lot of scientists who not only remember to think about DNA in this context, but also specifically study...plasmids containing these genes. As with the PIs above, many of these genes were discussed at the 2018 ...chromatin This is not the first Addgene blog post about chromatin--multiple Addgene colleagues and guest...

...pLionII 3rd FIV-derived (FELIX) plasmid with a chimeric 5’LTR, Any expression cassette (promoter and gene of... Read our Lentiviral Guide for more information about: Lentiviral components 2nd versus 3rd generation...systems Viral production Frequently asked questions about lentivirus use Glossary of commonly used lentiviral... or shRNA. See article for detailed information about cloning & additional plasmids. Aebischer and Trono...

...products are rare, probably accounting for less than 5% of repair events. However, the products of accurate...Break Diminishes Its Mutagenic Potential. PLoS Genet 5:e1000683 . https://doi.org/10.1371/journal.pgen.1000683...resolved during nonhomologous end joining. Nat Commun 5: . https://doi.org/10.1038/ncomms5286 Yan M-Y, Li ... extremely heterogeneous. There is consequently about a two-thirds chance of causing a frameshift mutation...favor accumulation of the latter products. As noted above, a single cycle of cleavage and accurate repair ...

...market; you need one with 5’->3’ polymerase activity (for amplification), 3’->5’ exonuclease activity (increases...(increases amplification fidelity), and no 5’->3’ exonuclease activity (which could potentially truncate...unique site in the coding region (preferably near the 5’-end) and that also generates 4 bp overhangs. You ... Brush up on Your Cloning Techniques Learn More about Plasmids in Our Plasmids 101 Series Use CRISPR to...