...for. Secondly, categorize your edit – is it any frameshift mutation? A specific insertion or deletion? A...line is desired, any mutation that introduces a frameshift in the coding region of interest will get the...and run the PCR product on a gel to look for size shifts. Once you’ve identified homozygous clones, however...large so that you can easily visualize the size shift you anticipate. A good rule of thumb is to keep ...

...initiation heavily dependent upon the core of a Shine-Delgarno sequence found at the 3’ end of the leader...was predictable based solely on the promoters and Shine-Delgarno sequences used in this so-called bicistronic...Academy of Sciences 113.9 (2016): 2388-2393. Huang, Shijie, Xuechen Zhu, and Charles E. Melançon III. "Detection...ChemBioChem 17.10 (2016): 895-899. Choudhury, Rajarshi, et al. "Engineering RNA endonucleases with customized...

...own piece (3). After incubation, blocking, and washing, the membrane is reassembled and imaged. In this...(PBS) are the most commonly used solutions for washing. If your background noise is high, you can consider...Like blocking, you don’t want to skimp on your washing steps! It’s important to let them wash for the ...sequence not accurate Check plasmid sequence for frameshifts and stop codons Weird background patterns...primary antibody at 4 °C overnight Incomplete washing Ensuring wash is happening under sufficient agitation...

...blue-shifted fluorescent protein (donor) and measuring the resultant emission from the red-shifted fluorescent...imaging. The solution is to couple Nluc with a red-shifted fluor. Here, NLuc internally excites the fluor ...these tools, NanoLuc-fluorescent protein fusion shifts NLuc’s bright blue luminescence to wavelenghts ...

...about fighting scientific partisanship, including more scientists in our leadership, and raising awareness...McLafferty is an editorial assistant and activist. Washington, DC, USA 38.9072°N 77.0369°W – Mark Lucera I...science education. Photo cred Alina Chan. Seattle, Washington, USA 47.6062°N 122.3321°W– Justin Sherman Prior...

...these plasmids for the majority of the countries we ship to, first on a temporary and now on a more lasting... exceptions for rabies and VSV-G genes, plasmids ship out approximately three weeks faster, as no license...addgene.org for the most-up-to-date information on shipping plasmids to your country. Curious about how rabies...

...monitor protein and cell ageing, as the spectrum shift occurs during Timer protein life due to a form of...read the Addgene interview with Mark Howarth. Shining a light on channelrhodopsins – Chronos & Chrimson...spectral peak at 590 nm, which is 45 nm more red-shifted compared to other variants. The group further optimized...functional GFP, and the cells will turn green. Mashiko et al., Sci Rep 2013 Nov 27; 3:3355. Rinehart...TALEN assembly and activity? The laboratory of Takashi Yamamoto has created a complete TALEN assembly ...

...and watch it now! (Bonus: Tim kindly offered to finishing answering all submitted questions via blog post... the next few weeks.)  Finally, Addgenies love sunshine and warm weather - check out our Instagram all...endeavors - in between some much-deserved vacations, sunshine, and beach trips, of course.    *not me, though...

... FACS machine after a single use. So how about washing and reusing pipette tips in a high-throughput and...example, offers automated, benchtop, high throughput washing devices that are compatible with a variety of pipette...incentives (for example, by grant agencies), we can shift toward a more sustainable one. The math on this ...

...of the method. One observation was an inverse relationship between deletion size and frequency [3]. There...coding sequences often result in loss-of-function frameshift mutations. However these may require restriction...genomic landscape may be surveyed in unprecedented fashion. Targeted deletions are just one of many available...

...muscular atrophy patients. Noushin Nabavi - Peritoneal Mesothelioma  Noushin Nabavi is a Postdoctoral Fellow...diagnostic tools and animals models of the disease in partnership with Taconic Biosciences. The study of peritoneal...

...repository in 2023.  In June, we announced a new partnership with the Institute for Protein Innovation (IPI...we’re pleased to announce that we are now able to ship rabies plasmids in our repository to international...through active conversation and collaboration.  Wishing you a healthy, productive, and thriving 2024, mirroring...

...Expanding your network of relationships early and often is the most effective tactic a scientist in training...result of existing professional (and personal) relationships. However, most job seekers will and should apply...after the academic bench. Utilizing existing relationships as part of the job application process can dramatically...

...person, and helps improve your interpersonal relationships with people you’re collaborating with remotely...like something out of your nightmares, shared ownership of materials produced from your collaboration ...the project. The state of research is constantly shifting, and long-distance collaborations are becoming...

...awesome. The process of writing, editing, and publishing blog posts runs its course more quickly than ...supervisors warned me, the swift gratification of publishing posts can be addicting. Even more satisfying,...always thoroughly enjoyed: travel. I’m actually finishing up this post on a plane back to Boston from Kentucky...

...deaminase from sea lamprey fused to Cas9 nickase (Nishida et al., 2016). Target-AID acts similarly but not...efficiency but higher indel formation than BE2 Nishida et al., 2016 Target-AID   Edits 3-5 base window...685. https://doi.org/10.1038/s41587-022-01533-6 Nishida, K., Arazoe, T., Yachie, N., Banno, S., Kakimoto...Mochizuki, M., Miyabe, A., Araki, M., Hara, K. Y., Shimatani, Z., & Kondo, A. (2016). Targeted nucleotide editing...

...whether I’m working with a talented journalist to shine light on living as a scientist in Beijing, talking...emotional investment and following cause-and-effect relationships are all things almost all of us are programmed...Interviewee: “I did the most work for a paper we were publishing, but when it came time to add author details ...

...new FP. Their aim - to red-shift the boundaries of FP technology by shifting the emission maxima toward...APOBEC-Cas9n-UGI-mediated activity edits out a frame-shift mutation within the mCherry reporter. Quantification...

...returned the MTA, Addgene can move forward with shipping the material to the requestor.  Why the MTA?  ...certainly frustrating to find out your plasmid hasn’t shipped because a form needs to be signed. But the truth... And that’s where MTAs, and similar agreements, shine.  These types of agreements allow for centralized...

... for clear communication, fair credit through authorship, and acceptance of a diversity of work styles...stress the fact that “miscommunication, a lack of leadership, and inappropriate tools or writing approaches...that was probably lost during these weird times of shift working and social distancing. The take home message...