We narrowed to 997 results for: lars

...CRISPR guide RNAs (gRNAs) for genome editing in large-scale has led to the rapid generation of genome-...combinatorially modify the genome, screen for a particular phenotype, and quickly profile the resultant ...Sciences of the University of Hong Kong. He is particularly interested in developing and applying new technologies...

... the gel in the first place. Carefully load a molecular weight ladder into the first lane of the gel. ...you are conducting certain procedures, such as molecular cloning, you will need to purify the DNA away ...you get better separation of bands? If you have similarly sized bands that are running too close together..., and a lower percentage gel will help separate larger bands. 10% Rule For each sample you want to load...

...cytosine (C), guanine (G), thymine (T)] in an equal molar ratio. It also requires two unique single stranded...copy the DNA in between the oligo primers. This molecular biology technique creates several micrograms of...Your 5’ and 3’ primers should be designed to have similar melting temperatures (Tm). Set the annealing temperature...frozen. DMSO helps with denaturation of the DNA, particularly for GC-rich templates. What does each ingredient...

...., 2020). Some Acr proteins are specific to a particular Cas protein while others can inhibit CRISPR enzymes...can increase the chance of off-target editing particularly after the majority of on-target editing has ...Cas9 Toxicity in Human Hematopoietic Stem Cells. Molecular Therapy - Methods & Clinical Development 9:390...

...sequence ranging from a single nucleotide change to large insertions like the addition of a fluorophore or...components. However, this sometimes requires intracellular injections into the cytoplasm or pronucleus ... albino mice ranging from 11% to 100%, a level similar to other approaches.  Nanoblades can also be used...

.... Sticky-end mediated gene transfer would be particularly helpful for targeting non-dividing cells, which... are 3.7 kb and 3.9 kb in length, respectively, larger than SaCas9 but smaller than SpCas9. These nucleases...et al. suggest that AsCpf1’s off-target rate is similar to that of high fidelity Cas9s eSpCas9 and SpCas9...

...point mutation or functional mutation in vivo: Similar to a conditional rescue experiment with a wildtype...sense (on) orientation in the presence of Cre. A similar strategy can be employed for "Cre-off" (i.e. your...for monitoring Cre-mediated recombination at the cellular level in the mouse. 2003 PubMed PMID: 12665802...

... parallel, this approach should be considered. Similar to the p24 assay, titering via measurement of viral...serial dilutions of a viral prep. This method is particularly useful when vectors carrying fluorescent markers...In-Depth Fluoresence Titering Protocol Another popular approach utilizes the antibiotic resistance genes...

...yourself anymore. Personally I found this to be particularly challenging. As PhD students and postdocs we...there is not a clear deadline. Finally check-in regularly, make yourself available, and encourage questions...manager for advice if you’re not sure how to face a particular situation. But keep in mind that a good manager...

...moist and hopefully aerated with a thin layer of molecular biology-grade agarose containing physiological... advantage of this technique is the potentially large number of single clones obtained (on occasion we...Maryland Medical School in Baltimore. She is particularly interested in signaling peptide maturation in...

...Cas9 technology has revolutionized the fields of molecular biology and bioengineering, as it has facilitated...gRNAs Importantly, the use of truncated guides has largely been demonstrated to result in decreased mismatch...technologies. To date his group has develop several popular CRISPR-based tools for altering DNA sequence or...

...see if any other fluorescent proteins display a similar effect (many do). Fusing EGFP to the self-labeling... tools, they’re optimistic that MagLOV can be similarly adaptable into future magnetogenetic tools.  Embracing...passive impact,” he says, noting that his team regularly shares troubleshooting tips and works with other...

...editor experimentalist, and especially shines once large-scale prime editing experiments come into play. ...Kissling, L., Marquart, K. F., Schmidheini, L., Solari, C., Balázs, Z., Krauthammer, M., & Schwank, G....DNA double-strand break repair pathway balance. Molecular Cell, 81(10), 2216-2230.e10. https://doi.org/10.1016...

...drug to the market usually takes over a billion dollars and at least a decade of work. The number of steps... Where publication is expected, the process is similar to academia. The only difference is that your paper...could be laid off while you’re there. Of course, similarly, an academic advisor could also lose their funding...

...cultures with intact ITRs. I use a spreadsheet similar to this one to plan my AAV packaging PEI transfections... This helps avoid multiple freeze/thaws of your larger aliquots of virus. AAV titration by qPCR Overview...in vitro and in vivo use. Current protocols in molecular biology, Chapter 16, Unit 16.25. PubMed PMID: ...

...figure below). The double-MMEJ strategy looks very similar to HDR, but it is accomplished using much smaller... of MMEJ. This double MMEJ strategy looks very similar to HDR, but it is completed using much shorter ...whole-plasmid integration if you’d like to integrate a larger amount of material into the genome. Open questions...

... being used. After learning of it's continued popularity, I decided this protocol would be a worthy first...Addgene has also cataloged a variety of useful molecular cloning protocols, including making LB agar plates...needs. At Addgene, we’ll continue to create new molecular biology educational resources, such as protocols...

...and reagents from other vendors should produce similar outcomes when using these protocols. However, please...blade Plastic tray Gel imaging system ImageJ or similar software Reagents Purified recombinant antibody...should be very little background staining. Pro-Tip A large shift in the electrophoretic mobility of the bands...Analysis Use ImageJ (Link opens in a new window) or a similar photo software to determine the relative intensity...

...Addgene, as a proxy for global activity in our particular scientific subfields.  Plasmid requests correlate...plasmid requests correspond to publications of popular papers. Figure 1: Plasmid requests by date...-examine our global data at a finer temporal granularity. For each paper, we observed that the demand ...

... Thermo Scientific NanoDrop is one of the most popular options, so most of this post is based on reading... from a NanoDrop. The principles will apply to similar instruments from other companies, like the Implen...reagents, making it more difficult to detect. A particularly strong absorber at 230 nm is guanidine thiocyanate...