...enriched pool of sybody candidates are then reverse transcribed into cDNA fragments that are efficiently subcloned...for binding to the target protein. The protocol describes going through two rounds of phage display to increase...interest. The Seeger lab used the ELISA technique described in their protocol because it is less susceptible...

...following cloning methodologies have been extensively described in other articles; so if you are interested in... to assemble up to 6 different fragments in a prescribed order. These fragments can be pooled in one tube...homology. You might also consider FastCloning, first described by Chaokun Li and colleagues. Here you simply ...

...template with the endogenous gene. Here, we'll describe the techniques and the plasmids used to inactivate...Conditional knockouts The methods and plasmids described in the preceding sections are simple ways of knocking...knockout, researchers often use the Cre/Lox system described earlier. In this case, you design your targeting...

...world examples of these situations in cells are described below (See Figure 1).   Figure 1: Each... not necessarily monomeric, even if they were described as monomeric in a publication. The gold standard... [2]. Some affinity assays used in the papers describing the FPs above including molecular sizing columns...

...made news in March with a publication in Science describing the development of “transcriptors,” transistor-like... and observed that we humans find it useful to describe and interact with the world using words like OR...The future of synthetic biology Addgene: You've described the pieces in these kits as the beginnings of ...

...from Alex Schier’s Lab. gRNAs can be in vitro transcribed from ssDNA, which can be generated by commercial...targeted approach using RNPs has recently been described in vitro, with the hope of eventually using this...PMC5681539. Additional resources on the Addgene Blog Subscribe to CRISPR blog posts Learn about other CRISPR ...

...structural proteins expressed from a reverse-transcribed piece of DNA called a “provirus.” This DNA is...proteins because they integrate this reverse-transcribed DNA into the host cell’s genome and then use ...is a turn of phrase — “pseudotransduction” — to describe the direct protein transfer of RT, which would...

...used in research, and data repositories. Below I describe how each of these kinds of connectivity facilitates...deposits will be made. The Dataverse integration is described in this video and the Figshare integration in ...from RSpace documents to external data sources (described above) further enhances the value the repository...

...restriction endonucleases in plasmid addition was first described by studies of EcoRI and Bsp6I in Escherichia coli...systems. As with the other toxin-antitoxin systems described above, one part of the pair is more stable than...toxin-antitoxin system discovered. Unlike the systems just described, the hok/sok systems consist of three components...

...designed, each binding to one strand of the reverse-transcribed target sequence that are present close to the...amplified by PCR using Taq Polymerase and then transcribed to provide RNA substrates for Cas13. Cas13, upon...fluorescence upon cleavage. Highlights: Reverse-transcribed RNA samples are amplified by PCR instead of RPA...

...genome editing with CRISPR/Cas9 in human cells was described in 2013 (Cong et al, Mai et al) we were very excited...thinking further about the distance relationship described above, we realized that, in addition to increasing... and neurovascular diseases. He worked on the described project while he was a Postdoc in Marc Tessier-Lavigne...

...site-specific incorporation of phosphoserine (as described below) provides researchers with a valuable tool...context-specific effects of UAG suppression, as described (6). To determine the stoichiometry of phospho-incorporated...phospho:non-phospho species on PhosTag gels as described above. Reagents and protocols Sep-OTSλ plasmid...

...promoters can be used in the repressible manner described above, as well as the inducible manner detailed...30 years since the original Tet-Off system was described by Gossen and Bujard, both the promoter and transactivator...cells, and can be used in the repressible manner described above, as well as the inducible manner detailed...

... been described in this blog. If your data does not meet the assumptions for the tests described above...error rate than the multiple comparison tests described above. Violated assumptions and statistics Statistical...

...inserted in this LTR so you don’t need Tat to transcribe the viral genome in producing cells. Currently...process is to test several protocols that have been described (protocol 1, protocol 2, protocol 3) to find the...quantify viral RNA. All of these methods have been described in a previous Addgene blog Once you know your ...

...Plasmid #52962) was a gift from Feng Zhang and is described in Improved vectors and genome-wide libraries ...was a gift from John Doench & David Root and is described in Rational design of highly active sgRNAs for...were gifts from John Doench & David Root and are described in Optimized sgRNA design to maximize activity...

...system is much more potent than the in vitro-transcribed single guide RNA (IVT sgRNA) approach (Dewari...comparison of Cas9 RNPs prepared with an in vitro-transcribed (IVT) sgRNA versus synthetic cr:tracrRNA. Cells...step by step protocol for the csRNP method is described in our recent article (Dewari et al. 2018), however...

...after the piñata burst, value was subjectively ascribed relative to the basic needs of each individual...There are 8 major categories found in the table, describing everything from basic to advanced functionality...Software design elements that fall into this category describe how the user is expected to interact with the ...

...in practice integration can occur. Okita et al describes a protocol for generating iPSCs by plasmid transfection...Methods RNA Delivery: For this approach, in vitro transcribed RNAs are serially transfected using a cationic...transduction, such as the polyarginine peptide described in Zhou et al and Kim et al. This is considered...

...an alternative method, called STARR-seq (self-transcribing active regulatory region sequencing) that tests...assay. In their recent publication, Kroeze et al. describe their enhanced TANGO arrestin recruitment assay...When used in combination with their previously described SpyTag/SpyCatcher pair, multiple protein fusions...to fuse the AID tag to endogenous proteins. As described in their recent Cell Reports paper, the Kanemaki...