We narrowed to 1,010 results for: RING;
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TypeBlog Post... first identified in the late 1960s during research at hot springs in Yellowstone National Park. In addition...Using these reagents and a series of heating (denaturing) and cooling (annealing) steps Taq polymerase... common for dNTPs and other reagents to run low during the final cycles which would slow or stop PCR amplification... as real time PCR is a PCR technique used for measuring a starting DNA concentration using PCR. qPCR requires...this fluorescent dye, the concentration of the DNA during the PCR reaction cycles is continuously detected... Variations Quantifying low abundance targets Monitoring subtle changes in target DNA levels Multiplex...
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Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity
TypeBlog Post...significant activity on some mismatch-containing pairings. During Golden Gate assembly, ligation of mismatched... assemblies that enable biosynthetic pathway engineering and optimization. However, current best practices...redesign was guided by both advances in the re-engineering of the original BsaI-HF® Type IIS restriction...efficiency and identification of mismatch-prone pairings. Using these 4 bp overhang observations, accurate...and a Development Group Leader overseeing a team bringing new products to market, New England Biolabs, Inc...comprehensive profiling of T4 DNA ligase fidelity and bias during DNA end-joining." Nucleic acids research (2018)... -
A Needle in a Base-Stack: Cas9 Structural Biology
TypeBlog Post...becoming an ever more ubiquitous tool for genome engineering, and studying its structure can help us understand...nucleotides 1-10 of the guide RNA, prearranged for base pairing with a complementary DNA strand. These 10 nucleotides...Figure 3 crystal structure, reflecting the less stringent recognition mechanism that can tolerate mismatches...mismatches in the next 8-10 bases after perfect pairing in the seed sequence. Figure 6: Electron...stabilizing the active conformation of the HNH domain, ensuring all binding partners are in place before cleavage...still being uncovered, and different techniques all bring their own advantages—and blind spots—to these questions...better understand how to manipulate it. Happy CRISPRing! Molecular graphics and analyses performed with... -
8 Great Depositor Research Stories
TypeBlog Post...you to our depositors for sharing your science with us! Corralling and Cornering the Goldilocks Antibody...editing and multiplex prime editing (requiring minimal payloads!) Uncovering Ancestral Bias in CRISPR Research...antibodies for use in mice and human brain IHC and sharing the data on the mAb3D atlas website. The iGluSnFR3...Beroukhim labs to write a post on their paper uncovering ancestral bias in CRISPR guide library design... -
Celebrating One Million Plasmids Shared!
TypeBlog Post...Reaching one million plasmids shared might have you wondering how many plasmids leave Addgene headquarters every...Joanne Kamens, Addgene’s executive director, said during our celebratory lunch, it’s hard to picture one... September 13, 2018 This massive scale reagent sharing led us to calculate how much time we have saved...would have saved 5 million hours through reagent sharing. How much is 5 million hours? A colossal 547 years...plasmids to the repository and for the scientific sharing community (check out the infographic below for ...the past month, we’ve enjoyed seeing scientists sharing their contributions on social media as well the... -
Lab to Office Culture Shock
TypeBlog Post...well. Making the transition from the bench can be jarring for the unprepared, and unfortunately, there aren...who referred to her lab as her family three times during a 10 minute conversation. Leaving a tight-knit ... “I’m a scientist.” What should you expect during the lab → office transition? I’ve already provided...sure, but it also eliminates that feeling of floundering one can have when starting in an academic lab...science,” and virtually every person you talked to during the day was also a scientist. But now you also ...upsetting, there are ways to get back to the bench during your off hours. DIY bio labs are cropping up in...organizations like Science from Scientists, which try to bring real scientists into schools to provide students... -
CRISPR-mediated Plant Base Editors
TypeBlog Post...editing in rice. In Genome Engineering: The CRISPR-Cas Revolution, Cold Spring Harbor Laboratory (2018)....et al., 2016) invented CRISPR-base editors by tethering cytosine deaminase with Cas9 nickase (nCas9). ...to an adenine base editor. A massive protein engineering effort by Gaudelli et al. resulted in the successful...it with dCas9, and transformed it into E. coli harboring the defective CamR gene. Screening for surviving...colonies and subsequent rounds of evolution and engineering gave rise to a mutant TadA (TadA*) that satisfactorily.... Kang, Beum-Chang, et al. "Precision genome engineering through adenine base editing in plants." Nature... -
Transferable Skills Guide: Career Planning Resources
TypeBlog Post... Career Planning Resources. The key to ensuring success during graduate school and beyond is to live in...transferable skills while working in the lab and by offering advice on areas where you might need some help...on what I have found to be especially beneficial during my own graduate career – and in fact, some of the... a job fair! Similar to a conference, job fairs bring you face-to-face with working professionals, with...the ACS career fair, which are larger meetings offering resources for students seeking employment within...graduate schools will even pay for you to attend!), exploring a job fair, and utilizing your local career development... -
Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research
TypeBlog Post...Transposon-Associated TNPB system. In Springer protocols handbooks/Springer protocols (pp. 245–260). https:/... genomic sites that can be targeted with TnpB, offering access to regions previously unreachable with ...infect cells, have emerged as promising tools for delivering these reagents in both plant and animal systems... enabling more precise and accessible genome engineering. More elaborate protocols are available in our...Additional Resources on the Addgene Blog CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9 RNA Interference... Additional Resources on addgene.org Genome Engineering Guide Plant Plasmids and Resources CRISPR Plasmids... -
Viral Vectors 101: Pseudotyping
TypeBlog Post...the host cell. Some of these envelope-receptor pairings are broad, allowing delivery into many cell types...allows these viruses to infect neurons. Besides altering the host tropism of your virus, there are many...reasons when you might pseudotype your virus: Lowering cytotoxicity. Different envelope proteins have... up less of the virus and is less affected. Altering sensitivity to serum. Different pseudotyped viruses...the TVA receptor in neurons using AAV and then delivering rabies dG pseudotyped with EnvA. This allows ...v12050513 Cronin J, Zhang X-Y, Reiser J (2005) Altering the Tropism of Lentiviral Vectors through Pseudotyping...Quick Guide to All Things Lentivirus Get tips for titering your lentiviral preps Resources on Addgene.org...