... RNA pool was cloned into bacterial expression vectors to create a library. Bacteria were transformed,...Jaffrey group was able to quantify the effect of mTOR inhibition on the activity of RNA Polymerase III...

... a variety of species, including the standard laboratory mouse. It’s also made targeted insertions relatively... mouse model. To avoid disrupting splice donor/acceptor sites, Miura et al. (2018) recommend targeting...

... a certain region or calculate a colocalization factor to assess the spatial relationship between the ...targets or you’re pushing the limits like Addgene depositor Ronald Germain (Radtke et al., 2020 multiplexed...

...Gurumurty et al., 2016). AAV can also be used as a vector to deliver CRISPR/Cas9 either through injection...Mediated Genome Editing with Adeno-Associated Viral Vector. iScience 9:286–297 . https://doi.org/10.1016/j.isci...

...expressing Cas9 via transduction with Cas9 lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some cells...gift from Feng Zhang and is described in Improved vectors and genome-wide libraries for CRISPR screening....

...PA-GFP. PA-GFP was derived from Jellyfish (Aequorea victoria) GFP (wtGFP) by mutating Threonine 203 to Histidine..."Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging." Nature methods...

...researchers think the enzyme needs cellular recycling factors to release its handiwork (Sternberg et al., 2014...conformational changes that make Cas9 such a valuable laboratory tool are still an area of active research. New...

...expected location of the target is an important factor in deciding which protocol is likely to work, we...more permanent knockdown, consider using shRNA vectors instead. With both siRNA and shRNA, it can be difficult...

...plasmid Cut vector - Background due to uncut vector Cut vector + Background due to vector re-circularization...ends on either side of the vector are compatible with each other, the vector is much more likely to ligate...fuse the two ends of the vector together). Protocol: Standard Insert + Vector DNA Ligation Before setting...amount of cut insert and vector to use for the ligation reaction. The volume of vector DNA and insert DNA used...where the insert is smaller than the vector) a 3 insert : 1 vector molar ratio will work just fine. We ...opens in a new window) ligation calculator to easily quantify how much vector and insert DNA to use. Combine... ALWAYS do a vector alone + ligase control. This will allow you to verify that the vector was completely...

...of this tutorial we will discuss how to add new restriction sites to the MCS of an empty vector. However...purify vector While waiting for your oligos to arrive, conduct a restriction digest of 1μg of vector with...Mix the annealed oligos with cut vector in molar ratios (vector:insert) between 4:3 and 1:6 in a standard...insert of 50bp in length into a 5kb vector, mix 100ng of the vector with 0.75-6 ng of annealed oligos)....just as easily to add short tags or shRNAs to any vector (the procedure will simply differ in terms of primer...primer design). Let's assume that your favorite vector has a relatively limited MCS (BamHI - EcoRI - SalI...restriction sites and add them to your existing vector. It is a couple days of work that will pay off ...