...complementary site in the genome where it will proceed to generate a blunt DNA double strand break. Cas9...stranded DNA degradation. Cas3 DNA digestion can proceed up to multiple kilobases, but only in one direction...) and does not require a tracrRNA (it can self-process pre-crRNA into crRNA). The size and simplicity ...11 cuts ssRNA and is capable of pre-crRNA self-processing. Functionally, it is very similar to Cas13 and...

...helps me see problem solving as a generalizable process.  Step One: Identify the problem and gather ...fix everything or revealed another issue in the process, you might need to go back to the beginning, reassess...strategy. Step Five: Share your results.  Does this process remind you of anything? Perhaps the Scientific ...be able to carry out the entire problem solving process on your own, but you can start by paying attention...

... at the mutation site, indicating that repair proceeded through NHEJ rather than HDR. Figure 1:...: Homology-directed repair in targeted zygotes proceeds from the maternal allele rather than the supplied... characterization studies, the edited embryos proceeded to the blastocyst stage at similar rates to untreated...surrounding such research, it is crucial that scientists proceed with caution and with proper oversight from regulatory...

...want to mentor you during the grant application process. When my PhD advisor first asked me to write a ...PI asks for assistance in the grant application process, she has faith in you! Additionally, keep in mind...motivating research will help you understand the process and continue to improve your grant writing skills...Still want more information on the application process? What types of writing feedback do you find most...

...staggering. The process is complicated. If you’re interested in the drug discovery process, or you want ...trials or has ties to industry, the drug discovery process can be a bit of a mystery to graduate students ...information. Where publication is expected, the process is similar to academia. The only difference is ...

...region of interest. Primers are crucial for this process as DNA polymerases require an existing strand of... Figure 1: Steps in PCR reaction   This process of denaturation, annealing and extension is repeated...programmed thermocycler. For more details on this procedure check out our PCR protocol page, protocol video...transcription-qPCR (RT-qPCR). The first step of this process requires RNA to be converted to cDNA using reverse...

...results we analyze as part of our quality control process has steadily grown. On a busy week, we may need...than 200 plasmids as part of our quality control process. Consequently our team has refined our use of the...might also enjoy seeing how our quality control process has changed with the introduction of next generation... reference/database sequence is ≥ 95%. [Our QC process would be trouble-free and much faster if 95% of...

...fewer of its own proteins, making the purification process simpler. Due to its high yields, E. coli is often...collect the cell culture supernatant for further processing. It may be helpful to incorporate regular feeds...also cautious during the aliquoting and storage process to prevent dropped tubes. Now that your antibody...Eriksson K, Shokri A, Chotteau V. Antibody capture process based on magnetic beads from very high cell density...

...target cells Labs tend to follow the same general procedure for lentiviral infections, produce the virus, ...vectors are engineered with non-native envelopes, a process called pseudotyping. Lentiviruses pseudotyped with...Segura, M.M., Kamen, A., Garnier, A. “Downstream processing of oncoretroviral and lentiviral gene therapy...cellular receptors for vesicular stomatitis virus.” Proceedings of the National Academy of Sciences 110.18 (2013...

...successfully navigated the trials and tribulations of the process, I’ve provided guidance and mentoring to several.... There are too many moving parts in the whole process to distill a one-size-fits-all recipe for success...Student? Get Tips on Getting the Most out of the Process Get more tips from the full eBook Of course there's...there's more to the job search and interview process – these are just my top takeaways. My full eBook (...

...Fluorescent RNA aptamers are used to track cellular processes by transforming cells with plasmids encoding aptamer-tagged...for qualitative and quantitative measurement of processes including RNA movement by tagging RNA sequences... Aptamer Fluorophores describes the enrichment process used to evaluate oligonucleotides for binding affinity...affinity to a specific target, and the molecular processes that make aptamer biology a powerful tool. In ...

...politics? Science is an apolitical process for seeking knowledge. The process begins with a testable hypothesis...hypotheses and the cycle begins again. No part of this process is inherently political so why is there a march...scientists as well as other citizens. Thus civilization proceeds in an informed fashion. For science to be useful...

...recording DNA abundance throughout the amplification process, rather than just at the end as in standard PCR...became the workhorses of molecular biology." Proceedings of the National Academy of Sciences 102.17 (2005...restriction endonuclease of Hemophilus influenzae." Proceedings of the National Academy of Sciences 68.12 (1971...sequencing with chain-terminating inhibitors." Proceedings of the National Academy of Sciences 74.12 (1977...

... menu of common cloning options in the deposit process in 2014 because of its gain in popularity. Gibson... the Miller Lab Protocol). The Gibson assembly process can be used to assemble up to 6 fragments in one...time commitment. Another advantage is that this process makes it easy to generate wild type and mutant ...drawback to the Gibson assembly technique is that the process works best with fragments over 200 nucleotides....

....   Overview of the restriction cloning process. Both the plasmid (blue, backbone) and the DNA ... beginning the restriction digest and ligation process, you should carefully choose your backbone and ...your experiments.     Alternatively, this whole process can be completed using a single enzyme if your ... more detail and verification required for the process to work well, so let’s go over the details step-by-step...

...workflow orchestration software for data pipeline processing (Nextflow), cloud infrastructure, and secure ...  In that post, we talked about Addgene’s data processing needs, our criteria for selecting a new workflow...single, organized pipeline and accelerated the processing of our sequencing data.”  Recently, Seqera published...

...division and large scale cell mobility are essential processes during embryonic development. These are required...differentiated body. In contrast, these dynamic processes are largely absent in adult organisms, where tissues... injury in the acutely inflamed glomerulus." Proceedings of the National Academy of Sciences (2016): 201606253...cell subsets in the skin-draining lymph node." Proceedings of the National Academy of Sciences 113.4 (2016...

...relaying key points about an unfamiliar topic or lab process. Along with our role as a plasmid repository, Addgene...-by-step instructions to navigate various lab processes.  Our most recent protocol video detailing Streaking...details every stage of the selection and checkout process on Addgene’s website: We’ll be continuously updating...

...viral vector. To start the AAV packaged on request process, simply navigate to your plasmid of interest. If...the same trusted production and quality control processes as our in-stock service: ddPCR, purity testing...GC/mL for AAV2. There are no quotes or special processes involved; simply request your plasmid of interest...

...the template and primer binding site, increase processivity, or improve thermostability. What came out on...increasing editing efficiencies 2-3-fold. However, the process of double nicking increases indel formation slightly...repair (MMR) complex, to temporarily inhibit this process and promote copying of the edited strand to the...to the numerous innovations that streamline the process of prime editing, the Liu lab contributed an improved...