... (a British idiom for “taking by surprise”) by Margaret Robinson’s lab, and more recently used by Stephen...Opto-RTK constructs – Opto-mFGFR1, Opto-hEGFR, and Opto-hRET – as well as various LOV-domain-mVenus constructs...directionally assemble in a single reaction, while retaining only a defined 4bp fusion site in between. The...vectors with the desired sequence of interest are retained after SLIC cloning. The general cloning strategy...

.... Pro-Tips: 1. It’s all about the MOI. In retrospect I used a way too high Cas9 MOI considering that...I didn’t do this, but to do it you would need a pretty good gel image. If you are analyzing short DNA ...cancer cell line in about 1 month, which I’d say is pretty amazing. Despite the shifting landscape of research...

... was not present (Kuhlman, SK. 2008). While theoretically this strategy should work, these vectors suffered...data and can instill greater confidence when interpreting the outcome.    References and resources References...Pillay S, Puschnik AS, Nagamine CM, Cheng F, Qiu J, Carette JE, Vandenberghe LH (2018) An Alternate Route for...

...gku241 Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P. D., Wu, X., Jiang, W.,...., Terns, M. P., White, M. F., Yakunin, A. F., Garrett, R. A., Van Der Oost, J., . . . Koonin, E. V. (...Pinilla-Redondo, R., Mayo-Muñoz, D., Russel, J., Garrett, R. A., Randau, L., Sørensen, S. J., & Shah, S....

...Off-target activities of the RNAi Correctly interpreting your data therefore necessitates careful design..., this can make the results more difficult to interpret, and a knockout/ or knockdown approach should ...validation is critical to ensuring accurate interpretation of your results. Therefore, taking advantage...

...Envelope protein for producing lentiviral and MuLV retroviral particles. Use in conjunction with a packaging...as pCMV-dR8.2 dvpr (lentiviral) or pUMVC (MuLV retroviral). Bob Weinberg pMD2.G VSV-G envelope expressing...

...proteins through mutagenesis. Chapter 6 - The future of FRET and fluorescent protein tools. Future developments...Find Fluorescent Protein Plasmids Find Plasmids for FRET Find Plasmids for Subcellular Localization ...

...drive change. This effort is already resulting in concrete actions and outcomes. For example, pd|hub assembled...collections of evidence-based practices. These concrete programs and practices to promote PhD professional...

...lungs are difficult, but other tissues like the retina have cell-based therapies underway. Chapter 7: ...Additional Resources from Addgene Learn about Retinal Gene Therapy from Connie Cepko Find Plasmids from...

...me to join the committee because I had recently retired from Harvard, and it let me keep my toe in the ...little bit?”    What are you doing to enjoy your retirement? I’m also on the advisory board for Art in Giving...

...toward a more sustainable one. The math on this is pretty simple: wash your tips once and you cut your tips...distribute it to industrial and residential markets. Rethinking our approach to lab waste is something we all...

...EBFP-Cre Available in serotypes AAV1, AAV5, and AAV retrograde (the most popular form), this viral vector is...with Flp-FRT. Additionally, you could use the retrograde form to specifically express Cre in neurons with...

...when all other options have been exhausted. Theoretically, phage alerts can be sent out to the directory... archive phage reagents. If a phage researcher retires, it's likely that the phages are lost if they are...

...foundation of multiple important technologies, including FRET and optogenetics. Even though GFP has been so thoroughly...characterized, it turns out this protein has a few more secrets - during a collaboration, members of Maureen Hanson...

...subtilis and the yeast Pichia pastoris. The improved secretory capacities of these microbes make them useful ...peptide (SP) that routes the protein through the secretory pathway to improve folding and disulfide bond ...

...New Hampshire in the summer for the annual lab retreat. How do you manage work-life balance? Tom Ellis... “three generations walk[ing] together in the Arboretum and other parks.” Lab life is not all work and...

...for packaging: AAV1, AAV2, AAV5, AAV8, or AAV Retrograde (AAVrg) capsids.  Please be aware that this ordering...plasmids packaged in AAV1, AAV5, AAV8, and AAV Retrograde to be 1x1013 and titers for AAV2 to be 7x1012...

...for the performance of a reagent, e.g., antigen retrieval conditions, dye inactivation details, tissue preservation...preservation, target tissue, detergent, antigen retrieval conditions, and more. Users can also recommend...

...ahold of your destinies” and also suggested some concrete actions that can be taken now to solve some of...Disequilibrium, Disillusion and The Postdoc Dilemma” which pretty much said it all.  She cited the oft quoted statistic...

...speechless (the club of people who have been in space is pretty small). Therefore, I was delighted to be invited...During the panel, an artist was sketching her interpretation of the discussion in markers on a whiteboard...