... be stable in the cell. gDNA guides will also need to be transfected into the cells of interest. In contrast...unable to show TtAgo-mediated cleavage in mammalian cells, likely due to high temperature requirements. To...plasmid and genomic DNA in a variety of mammalian cell lines, with an efficiency similar to that of Cas9...

...planning for your visa and or green card process well before addressing your career alternatives in order...the United States including green cards that are well-tailored to meet the needs of researchers. The categories...related skills”. Many career paths in academia as well as the non-profit sector are transferrable to industry...national importance; (2) The foreign national must be well positioned to advance the proposed endeavor; (3)...below. An international or national award for excellence in your field. Membership in a professional organization...

...spend the afternoon lovingly tending to the stem cells I was using in an experiment, then spend the evening...evening listen to some bloke at the pub telling me that actually he drives better after a few drinks. A...it’s oddly fitting that I’ve ended up (again) dispelling the same myths and misconceptions fifteen years...Kahan, D., Peters, E., Wittlin, M., Slovic, P., Ouellette, L. L., Braman, D., & Mandel, G. (2012). The polarizing...

...CRISPR to Cats Stem Cell Models for Disease & Open Science: Interview with Darrell Kotton Learn How Plasmid...while some are looking to develop new skill sets. Kellen Andrilenas is a Ph.D. grad student at Boston University...engineering project at BosLab. Once he optimizes plant cell culture and transgenesis he wants to share his knowledge...

... colleagues. When you attend a well-run meeting, note what worked well and ask the meeting leader for ...annoying interruptions to our “real” work. However, a well-run meeting can have quite the opposite effect. .... We’ve provided some tips above for what works well at Addgene, but meetings are never one-size-fits...

...expected to make sure the work gets done and done well. Involving your team effectively is now your new... in mind, you won’t be able to communicate these well to the task assignee and this is a recipe for bad...delegating is a bad idea. How do I make sure it goes well? As with most managerial responsibilities, good ...Delegating" in the Harvard Business Review. Another excellent blog on delegation tips and practices can be found...

... R, Schmalhorst PS, Jude JG, Zuber J, Gerlich DW Cell. 2017 Aug 24;170(5):956-972.e23 Screen identifies... R, Schmalhorst PS, Jude JG, Zuber J, Gerlich DW Cell. 2017 Aug 24;170(5):956-972.e23 Rapid and phosphoinositol-dependent... Rando OJ, Xue Y, Swiderek K, Kuo A, Crabtree GR Cell. 1998 Nov 25. 95(5):625-36. An architectural role..., Ensminger AW, Fox AH, Chess A, Lawrence JB Mol Cell. 2009 Mar 27;33(6):717-26. Reporter-nanobody fusions...

... be wondering how this all fits in with science. Well, I’ll give you an example. At my previous company... goes into software product management? I always tell people the best part of my job is getting to talk...communicate with customers, software developers, as well as team members in automation, sales, marketing,...help you be a valuable asset to the scientists as well as the developers. Lastly, you’re going to need ...

...idea from academia to a startup Mike Koeris and fellow co-founders Tim Lu and others first came up with...non-scientific audiences. "There are 100 ways to tell a story - convince your audience you are doing amazing...aligned in vision and outcomes. The team must work well together, have complementary skill sets, and “talk...were free and free to repair.” If the team works well, the idea is solid, and “the pitch is packaged to...

...you correct any language- and spelling mistakes. A manuscript that is well written looks more professional...submission, missing files or a manuscript full of spelling errors will not have a great chance of getting...manuscripts submitted for publication. Holds a PhD in Cell Biology and a Masters degree in Infection and Immunity...

...Facebook and Gmail, were launched. And Melina Fan, a cell biology graduate student, was frustrated by the ...to receive the Massachusetts Nonprofit Network Excellence Award for our electronic MTA system. We also ... our most popular lentiviral and AAV vectors, as well as developing and sharing supportive educational...stories we can, and to hear the stories others have to tell.  "Also thanks for being Addgene. You guys rock....

...enable scientific reproducibility. This workshop was well received, and then when the team started brainstorming...widespread issue of irreproducibility. Use your network well to seek out valuable and impactful opportunities...course! Thanks to individuals in the group who were well connected, this diverse and international team assembled...website How-to-guide for Reproducibility workshops Levelling Up: Reproducibility for Everyone Addgene: a global...

... channels in living cells. Multiple properties of a channelrhodopsin, such as cell localization, kinetics...would result in a functional channelrhodopsin, as well as predict the channelrhodopsin’s photocurrent strength...minimally-invasive optogenetics While ChRgers performed well in a tissue culture dish, that doesn’t guarantee...

...practical solutions.  My PhD prepared me remarkably well for this task. Good scientists learn how to communicate...to say “I’m going to pour these plates now” to a fellow team member. But, at Addgene, we use seven different...different types of “plates” (Fig. 1). When you have to tell an engineer that you want the LIMS software to create...It is never really done. But if you implement it well, here’s what can happen: More effective meetings...

...read a 100-page genre fiction novella as it does to read an 11-page Cell paper - and sometimes, it might...obtain?” If the answer is yes, you will need to spell out that connection for your audience. I ask myself...needed. The trick is to communicate your key points well enough that the audience can take that knowledge...

... project aimed at understanding how a critical cellular pathway is regulated. You’ve read all the background...generated against a linear antigen tend to work very well for applications that denature a protein to its ...expression in knock-out or knock-down down tissues and cell lines; orthogonal strategies that compare antibody...References Uhlen M, Bandrowski A, Carr S, Edwards A, Ellenberg J, Lundberg E, Rimm DL, Rodriguez H, Hiltke T,...

...teams continue to keep up with the demand with excellence and dedication. We don’t have worry or wonder...right? Upgrades to the internal LIMS pages and excellent work on our external website appearance and functionality...support. The Addgene management team leads with excellence. They are one of the main reasons I have so few...Addgenies. I never worry about their dedication, excellent intentions, or incredible drive to succeed.  Thanks...

...transfer 100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see image above)....integrates our database of over 2000 primers. Two 96-well PCR plates are prepared with the primer stocks for... be found here. Finally, you will want to have a well-organized database of expected sequences, either...some tips on how to get better sequencing reads? Tell us your favorite sequencing tricks in the comments...

... Fragmid website, or through Addgene, has worked well in the hands of the Doench lab. Additionally, controls...Drosophilia Plasmid pRDB_051 (for stable fly cell lines; attL and attR sequences for genome integration...Drosophilia Plasmid pRDB_052 (for stable fly cell lines; Hygromycin resistance gene; for genome integration...rapid assessment of emerging CRISPR technologies. Cell Genomics, 4(3), 100519. https://doi.org/10.1016/...

...colony.  You should genotype all of your F1 mice, as well as progeny from future generations. 4. Continue ...strain, the promoter being used should be tested in cell lines to determine how it impacts the expression...on separate chromosomes in future generations as well, a concept known as linkage. This hinders the likelihood...interest. Cre may be expressed globally (in every cell of the mouse) or in a tissue-specific manner (with...

...communication, and policy. In this post, I hope to tell you a little more about how I ended up at pgEd, ...starting conversations about both the benefits as well as the personal, social and ethical implications...broaden my training beyond the biological sciences, as well as to acquire leadership and communication experience...scientific, educational or industry conferences, as well as schools, community events, churches or mosques...

...answering help emails and phone calls, I feel compelled to share my years of accumulated wisdom so that...company’s FAQs? A lot of companies and organizations selling biological reagents have a ton of data on the catalog...their products from you, unless they’re knowingly selling a bad product (in which case, you should probably... some time. If the customer service rep does not tell you upfront how long it will take to get an answer...

...uses a small molecule, a tag, and the endogenous cellular degradation machinery to function. The dTAG is...tag and small molecule. It does work in mammalian cells, mice, and even yeast, though (we have a kit!). ...and complete degradation of proteins in mammalian cells. Proc Natl Acad Sci U S A. 2012 Dec 4;109(49):E3350...Tags Plasmids for Endogenous Gene Tagging in Human Cells Antibodies 101: Epitope Tags  ...

... thought about management skills before being propelled into the position of manager. I barely had any...check myself sometimes to make sure I am delegating well, but it does get easier over time! Communicate often...to recognize someone’s hard work and reward a job well done. Your report will (needs to) feel valued. Talk...should help you work through your situation, not tell you what to do. Unfortunately this is not always...

... on how your virus of interest causes cancer, as well as therapeutic strategies to target it, than you...at an unfamiliar venue. Finding your poster board well ahead of time ensures you aren’t scrambling to tack...published papers or “hot topics,” so be sure to be well-read on the recent literature in your field. Take...know every answer – don’t hesitate to be honest and tell your audience just that. Explain what you do know...

...is an excellent resource that has gained a lot of traction with researchers. It is an excellent way to... future career. If it wasn’t something I liked – well, I was used to experiments not working, so just ...programs and departments Graduate school was an excellent time to try new things, develop new skills, and...

...his own biases--many of which will be shared by fellow scientists--but that doesn’t make his introduction...have outgoing personalities to supplement their intellectual prowess and scientific skills. This is an era...has been such a great fit for me!), the book was a well-executed overview that recontextualized some areas...career.  Acknowledgements: I would like to thank fellow Addgenies Joanne Kamens, Marcy Patrick, Aliyah ...

...transformation efficiency of the strain: when asking the cell to perform multiple recombination events, all linear...linear fragments need to get into the same cell. For simple cloning requiring just one or two recombination...recombination events, homemade competent cells of 107 CFU/μg are perfectly fine, yet performing multiple modifications...modifications will require ultracompetent (109 CFU/μg) cells. Primer design Using specific primer design (Figure...

...using an electric charge to help the DNA enter the cells. Some samples require a reduced antibiotic concentration...concentration or need to be transformed into different cells than were originally requested. Most of our samples... Start by scanning the sample barcodes into the cells in the same order that they exist within the rack...storage process - from deposited DNA to transformed cells and finally to bacterial glycerol stocks that are...

... be safely used to deliver a genetic payload to cells, and the complexity of effectively deploying them... can be to use. AAVs, which can target different cell types depending on their serotype, are particularly...amount of vectors available and number of requests tell only part of the story. Viral vectors are, after...experimental data in a standardized format, sharing how well a given viral vector worked — or didn’t work — in...