...This post was contributed by Kartik Lakshmi Rallapalli, a graduate student at the University of California...manipulate it. The term genetic engineering itself is attributed to SciFi writer Jack Williamson who used it in... Zinc finger nucleases (ZFNs) are artificial restriction enzymes generated by fusing a zinc finger DNA-binding...template.  Each zinc finger recognizes a nucleotide triplet and when several ZFNs are strung together, it can...the ZFNs, but instead of recognizing nucleotide triplets, the components recognize individual nucleotides...

...This post was contributed by guest blogger Søren Hough, a Biochemistry PhD candidate at the University...mismatch along the length of the gRNA given the stricter sequence requirement at the terminal 3’ PAM site... Dillard, and Ayokunmi Ajetunmobi for their contributions to this work. Søren Hough is a Biochemistry ... mismatch cleavage assays to detect mutations triggered by engineered nucleases." G3: Genes| Genomes| ...PMID: 27087594. 16. Tycko, Josh, Vic E. Myer, and Patrick D. Hsu. "Methods for Optimizing CRISPR-Cas9 Genome...

...scientists at 5,200 institutions in 82 different countries. We continue to ship out more plasmids each year...beginnings in 2004 to today, you can see how plasmid distribution has grown. Additionally, more and more scientists...research. *2015 total reflects the number of distributed plasmids as of March 26, 2015. How does Addgene... our depositors to allow us to help store and distribute their plasmids. The partnerships and collaborations...

...SaCas9 genome editing plasmids String assembly gRNA cloning (STAgR) String assembly gRNA cloning (STAgR)...studied Cas12b nuclease from Alicyclobacillus acidoterrestris (AacCas12b) exhibits optimal DNA cleavage at...STAgR), developed by Stefan Stricker’s lab, is a single step gRNA multiplexing system that generates vectors...

...component of Golden Gate and GreenGate are type IIS restriction endonucleases. They are like other endonucleases... should in principle work in any plant species. Tricks of the trade Lohmann does have one special tip ...their own lab: be very careful with the type IIS restriction endonuclease of choice, BsaI, which can be very... will get busy with GreenGate, producing and contributing more modules into the public domain in the process...

...This post was contributed by Doug Richardson, Director of the Harvard Center for Biological Imaging and...focus fluorescence from reaching a detector or restricting fluorescence to specific excitation volumes to...reduction in signal intensity. 2-Photon microscopes restrict excitation (and bleaching) to a single focal point...microscopy uses a number of optical and chemical ‘tricks’ to turn subsets of fluorescent molecules on and... a large axial range. 2-photon imaging, which restricts excitation and photobleaching to the focal plane...

...detergents such as triton-x, saponin, or tween. Take care when using strong detergents like triton-x and tween...fluidics system they pass through a flow cell which restricts the size of the stream and forces the cells to...cell expressing low levels of the protein. The electrical system converts the fluorescence intensity to...you would need a way to determine the relative contribution of yellow and orange from FITC and that from...

...cells that have been infected with a microbe, to trigger immune cascades that clear these infections. Anything...letters and be able to guess whether or not that string of letters is a part of the word you’re searching... that you want the antibody to recognize. This triggers an immune response within the animal, and its ...antibodies (i.e., immunofluorescence), or by a colorimetric reaction facilitated by an enzymatic reaction... antibodies for neuroscience research from the Trimmer lab ...

...various grades of plastic lumber and distribute it to industrial and residential markets. Rethinking our...this, we can use the ACT label, which is an eco-nutrition label for laboratory products that emphasizes ...typically is destined for a landfill. However, Triumvirate Environmental developed Red2Green, a program ...

...in the same vector backbone with very similar restriction enzymes. You can essentially cut and paste quite...mind that a very simple sequential or double restriction enzyme digest will open up possibilities exponentially...these vectors to the scientific community and contribute in a way that everyone could utilize what we ...really the one who had been very adamant that we distribute to as many researchers as we could, and Addgene...

...programs either. But with a little Googling and some trial and error, I found some free design software that...number of drawing tools, so it’s harder to draw intricate figures, like a brain or a mouse. Vectr Vectr ...and digital articles where it’s easier to give attribution, than for journal publications. Google Image ...includes images that can’t be reused without attribution, despite filtering for images labeled for reuse...

... al., 2020). A transducer complex consists of a trimer of three subunits: alpha, beta, and gamma (Gα, ..., Gβ, and Gγ), collectively known as a heterotrimeric G protein. This complex converts a signal received.... When inactive, these subunits are found in a trimeric complex bound to the intracellular surface of ...called BRET2. Figure 1: TRUPATH measures heterotrimeric G protein dissociation by bioluminescence resonance...

...let us know in the comments. Note:Marcy Patrick contributed to the writing of this article. ... Short segment of DNA which contains several restriction sites allowing for the easy insertion of DNA....utilize one of a variety of cloning methods (restriction enzyme, ligation independent, Gateway, Gibson...

...fulfill our mission by generating revenue through distribution of research materials. Our small shipping and...generate revenue (as Addgene does through materials distribution). In fact, healthy nonprofits should have some...their revenue. A nonprofit organization cannot distribute profits to private individuals through stock ...request plasmids (via web interface, processes, distribution, and customer service) Addgene provides unmatched...

...the first gRNA using restriction enzyme BbsI and the second gRNA using restriction enzyme BsaI. If you’... of gRNAs Golden Gate Assembly uses Type IIS restriction enzymes, which cleave outside of their recognition...These expression vectors each contain Type IIS restriction sites flanking the promoter-gRNA construct, but...oligonucleotides. These plasmids contain Type IIS restriction sites that flank the promoter-gRNA constructs...

...This post was contributed by guest blogger Jon Backstrom, a biochemist in the Vanderbilt Eye Institute... have been immobilized on a reduced glutathione matrix. The advantage of a fusion protein is the efficient...provide a more relevant kinetic analysis, but the trick is to utilize a second immobilization method. One...available. Factor Xa loves calcium, but metal-chelate matrices, not so much. Maybe a calcium-independent proteinase...

...! Archon1, a voltage sensor AAV Historically, electrical activity in the brain was measured with implanted...voltage reporter allows a way to monitor neuronal electrical activity through imaging. Since it’s illuminated...particular cell types. For example, the mDlx enhancer restricts expression to GABAergic interneurons. These vectors...pAAV-hSyn-EGFP *Also coming in our new 20 ul trial size! Voltage indicators Plasmid Serotype Name...

...protein to regulate cell growth dynamics Article contributed by Klaus Wanisch  Listen to the optogenetics ...visualizing RNA production and localization Article contributed by Tyler Ford  Listen to the riboglow podcast...developing new FPs - the mCarmine story Article contributed by Caroline LaManna  Listen to the mCarmine podcast...base-editing and Cas9 cleavage activity Article contributed by Angela Kaczmarczyk  Listen to the base editing...

...The triple T’s of biology are transformations, transfections, and transductions! In this blog we will...basics but are here because you want to know the intricacies of transductions, we've got you covered there... easy. Producer cell media can be collected, centrifuged to remove any producer cells floating in it, ...long-term (AAV). If you perform a lipid-based or electrical-based transfection, most of the nucleic acid ...

...training progresses.  In From Academic Solos to Industrial Symphonies, an article well worth reading on ... notice or choice Always working on a team/in a matrix - can be frustrating for solo flyers Other people...—science can be surprisingly more rigorous in industrial settings Tight (sometimes impossible) timelines—others... years—a short and exciting ride Complicated matrix structures are common Small integrated teams, ...