...fluorescent reporter assays to quantify targeted Cas9 cleavage, and repair through homology directed repair (...while cells containing the reporter gene that was cleaved by Cas9 and repaired via HDR using the supplied... seamlessly remove the integrated GFP reporter leaving you with just your desired genomic edit.  Find ...using GEmCherry2. In this experiment they saw a clear difference in mCherry expression using FACs sorting...

...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...

...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, ...

...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...

...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...

....) 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...

...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...

...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...

...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-...

...Addgene blog Learn about other CRISPR methods for bacterial genome engineering   Learn about human CRISPRko...environmental isolates. For example, Peters and colleagues used Mobile-CRISPRi to knockdown individual genes...-CRISPRi. As a proof of principle, Peters and colleagues constructed a 40-member library targeting 20 ...

... 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...

...Arabidopsis in Education Learning Synthetic Biology with BioBuilder 3D Models for learning about CRISPR Resources... subsequently destroyed by Argonaute-catalyzed nuclease activity or their translation disrupted through...protocols and want to share pictures, feedback etc. please write to [email protected] and if you make...

...Addgene Blog Learn about Synthetic Biology with BioBuilder Use Addgene Videos to Learn about Basic Lab...genes in other species, but these studies generally leave questions about gene function wide open. What if...Check Out Our Molecular Biology Reference Pages Learn Simple Lab Techniques in Our Protocol Videos ...

...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...

...spectrum. The experimental apparatus should be cleaned thoroughly between animals as olfactory stimuli...animal (ex: urine) can be distracting. If it is not cleaned, animals tested towards the end of an experiment...coordination to more complex situations such as learning and memory or emotional states, for example anxiety...Guide to Viral Vectors Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

...unfortunate experience of witnessing…nothing. For some unclear reason, cells are not getting transduced. Here,...reducing the culture volume into which the virus is released, a more concentrated virus solution can be obtained...PMC330758. Additional Resources on the Addgene Blog Learn All about Adenovirus Associated Virus New to Virus...Virus? Check out Our Primer for Beginners! Learn All about Lentiviral Vectors Additional Resources on ...

...adenovirus, and adeno-associated virus (AAV). The least risky of these four viruses is AAV – it is not known...propensity to integrate into proto-oncogenes, which can lead to cancer. These viruses need to be handled in a...thoroughly decontaminated after use, typically with bleach (plastic) or ethanol (other surfaces). For AAVs...and isopropyl alcohol, 0.5% peracetic acid, 10% bleach, or 1% iodine are useful. Some organizations or...