...collection in your experiments and published your data, please fill out this form and we’ll add your citation ...combinations of WT, codon optimized, untagged, cleavage sites (thrombin and TEV), and C-terminal tags ...downstream experiments and offer flexibility in tag, cleavage, codon usage, and expression system. Mammalian...Addgene blog Read all of our COVID-19 blog posts Learn about other COVID-19 plasmids available at Addgene...

...stakeholders to learn more about their needs and any problems that they are trying to solve. After learning more ...Technologies, I work with our customers in the US to learn where the gaps are in their sequencing workflows...problem-solver. A lot of the time, the problem to solve is clear, but the solution is more complicated. You’ll need...

...Addgene blog Learn about other CRISPR methods for bacterial genome engineering   Learn more about lambda... novel genetic variants in bacteria. Read on to learn about the key components of EvolvR and its potential...extends it from the 3’ end, while its native endonuclease activity degrades the displaced strand.   ...

...ever like to write about a recent plasmid deposit please sign up here.  Listen to the hot plasmids episode...Vellai lab has created an improved RNAi vector that leads to significantly stronger phenotypes than standard...incorporation of non-specific nucleotides into the dsRNA leading to increased RNAi efficiency. Find the T444T ... forms an active LacZ tetramer.  This tetramer cleaves chlorophenol red-b-D-galactopyranoside on the assay...

...experiments. As a common example, a selection will leave you with the colonies that contain your plasmid ...negative controls for your PCR.  Find the details and learn more about colony PCR in this blog post.   ...can confirm the sequence of the entire plasmid. Please note that NGS verification is more time-intensive...biology protocols and tips Resources on Addgene.org Learn more about Sanger sequencing Read the Molecular ...

...ever like to write about a recent plasmid deposit please sign up here. Here's what you'll find in this...a continuation of that work, they have found at least two variants that display increased neuronal expression...microbial SSAPs (dCas9-SSAP) which insert genes in a cleavage free manner. The dCas9-SSAP system reports an ... less risky.   Fig. 3: Development of a cleavage-free dCas9-based gene editor using microbial SSAPs...

...ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll find in this...of anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-...anti-FLAG-FB-GFP labeling various targets with nuclear (top, 4x-FLAG-mCherry-H2B), cytoskeletal (middle, 4xFLAG-mCherry-β-actin... editing, while gene editing can be done via a nuclease-active CasMINI. This small yet powerful system...

...option, in aliquots no smaller than 10uL. You can leave them at room temperature when you’re working with...illustrious lab career, you will likely accidentally leave your antibody out on the bench overnight. Do not...this post, the conjugates, or signaling molecules! Please keep in mind that this is just a brief introduction...data, you’ll want to pick fluorophores with the least spectral overlap between them whenever possible....

...the nucleus through nuclear pores and thus do not require breakdown of the nuclear envelope during cell...there – most of which we would all like to steer clear of! However, a subset of viruses are commonly used...which ones could be helpful to you. Interested in learning more about a specific virus? We have you covered...

...Additional Resources on the Addgene Blog Learn More About Lentivirus Learn more About AAV Get the Scoop on Our...of 2016. The top viruses of 2016 were (drumroll please)... Top Lentivirus: lentiCas9-Blast Most of Addgene... to perform unique editing tasks, classic Cas9 nuclease, which introduces double stranded DNA breaks, ...

...β-galactosidase mutation. An unsuccessful cloning reaction leaves the α-peptide intact, and therefore the cell will...artifact that disrupts the α-peptide DNA will also lead to a white colony. And also false negatives: These... fragment is inserted in-frame, read-through can lead to a functional β-galactosidase enzyme and a blue...Blog Read Our Post on Common Lab E. coli Strains Learn to Use Yeast Two Hybrid Systems Browse All of Our...

... suited for particular applications. Read on to learn more about each reporter or check out table 1 for...nanobody that would make a useful RANbody, consider leaving a link to its sequence in the comments below. ...29440485. Additional Resources on the Addgene Blog Learn more about using the secondary nanobody toolbox ... plasmids can be found here Watch this video to learn more about nanobodies Additional Resources Looking...

...This biosensor changes fluorescence based on the release of glutamate from neurons and this new version ...than its predecessors. Currently, Aggarwal, and colleagues Filip Tomaska and Tim Hanson, are characterizing...iGluSnFR3 in vivo and have used it to detect glutamate release from hundreds of synapses at once. “This work has...Aggarwal.In addition to the biosensors, Aggarwal and colleagues are working on developing new surface display...

....) Fab fragments are created through enzymatic cleavage of the variable region (the antibody region that...to the antigen) from existing antibodies. Each cleaved antibody will produce two ~50kD Fab fragments, ...fragment but created through engineering instead of cleavage. They are comprised of the variable heavy and ...Whatever affinity reagents you use, one thing is clear: it’s an exciting time to be a scientist looking...

...week could expect to see their salary raised to at least a new legal minimum of $47,476 per year, under updates...universities with the FLSA in our online resource leading up to this date, as discussed in a previous Addgene...planning unionization at their institutions. One clear message that institutions have sent, with the lack...the Advocating for Science Symposium and Workshop Learn about a Variety of Science Careers Resources on...

...does it play in modern cloning, and why should you learn more about it? Read on to find out how ccdB can ...restriction enzyme-mediated disruption of ccdB, leading to positive identification of desired clones. ...most appropriate method for your own experiments. Please check out Addgene’s Choosing a Molecular Cloning...CcdB protein induces efficient ATP-dependent DNA cleavage by gyrase.” J Mol Biol. 1993 Dec 5;234(3):534-...

...Troubleshooting DNA Sequence Learn Additional Ways to Verify Your Plasmid Learn How Addgene Uses Barcodes.... All of our incoming plasmids are sequenced at least twice, and sometimes more depending on the number...read. You can expect to recover around 750 bp of clean sequence from a typical reaction, or about 250 amino...

...on the symposium as a whole and for registration please go to http://futureofresearch.org/boston/. The ... data and the Labor Market. On this topic it is clear that many important questions, areas for informed...biomedical_research_wgreport.pdf Resources at Addgene Learn More about Careers in Science in Our Career Posts...Meetings from Addgene Executive Director Joanne Kamens Learn how to be a successful manager with our Management...

... "traditional" restriction enzymes in that they cleave outside of their recognition sequence, creating... BbsI), that are located on the outside of the cleavage site. As a result, these sites are eliminated ...a few advantages over other cloning methods. Exonuclease-based methods like Gibson assembly require 20...multiple DNA fragments to create TAL effector nucleases. These plasmids were designed to utilize the BsaI...

...the promoter region of gamma hemoglobin (HGB1/2) leads to continuous expression of fetal hemoglobin even...efficiency, respectively) and outperformed ABE7.10 by at least 1.4 fold. The ability of ABE8.20-m to edit primary...editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464–471 . https://doi.org/10.1038/...Addgene blog Find more blog posts about base editing Learn about another evolved base editor: ABE8e Read about...