...resistance gene (missing the first two codons). Two stop codons are inserted between the truncated gene and...ligated into the MCS/SacI site in pUCXKT, removing the stop codons during backbone digestion and providing the...Toolkit for constructing fusion proteins with customizable N- and C-terminal modules   Thanks to the lab...Technologies. The MultiSite Gateway Kit is a customizable Gateway®-based toolkit that allows scientists...bound to the mRNA since they are tethered AFTER the stop codon. Thus a non-translated mRNA will appear yellow...: 10.1126/science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector...LacZ, puromycin or CreERT2 can be combined with custom-cloned vectors in a Multisite Gateway reaction ...

...Transformation Plasmid Cloning Creating Bacterial Glycerol Stocks Restriction Digests Guides Browse our in-depth ...

...IP and ChIP assays. Tag 96 genes at a time using Custom Synthetic cr:tracrRNA/Cas9 Ribonucleoprotein (csRNP...first step of optimization, we started off by using custom synthetic two-part cr/tracrRNAs for CRISPR experiments...tagging, pick a crRNA that cuts 8-15 bp downstream of Stop codon; for N-terminus tagging, a crRNA cutting 8...csRNP experiments, it can be ordered in bulk and stored in small aliquots at -80 oC. Assemble csRNP: Anneal...near the insertion site (within ~8-15 bp from the stop codon for C-terminus tagging) GC content of the ... hands, delivering Cas9 protein in complex with custom synthetic cr:tracrRNAs (csRNP) is the most efficient...

...not necessarily represent the views of Addgene. Boston, MA, USA 42.3601°N 71.0589°W – Alina Chan The ...dabbles in science policy, public outreach, and restorative justice.  Follow her on twitter @sung_vanessa...to join the public conversation and to tell the stories behind our research.  That's why I marched for ...Michigan, where programs like the Great Lakes Restoration Initiative are at risk of major cuts and where...behind these two fingers the ninety-nine cubits of Aristotle, nor would you mention my small error and at the...second and that it is our duty to tell our science stories in a manner that encourages open discourse with...

...Emission Depletion (STED), and Stochastic Optical Reconstruction Microscopy (STORM). These dyes have specific...Throughout their history, fluorescent dyes have enabled the visualization of both organic tissues and...than was absorbed. This phenomenon is known as the Stokes Shift, or the difference in wavelength between ...technologies in fluorescent dyes/ligands Throughout the history of cell biology, fluorescent labeling techniques...fluorescent proteinFluorescent Proteins 101: A History of Fluorescent Proteins  ...

...skeptical reader of historical fiction (I can personally attest to this!). Following history from the Ice Age...Wrong - and the New Research That's Rewriting the Story by Angela Saini "This book is a smart and carefully...Strange & Mr. Norrell by Susanna Clarke "A quasi-historical/fantasy fiction. Two dueling magicians in the...of England by Edward Rutherfurd "A 10,000 year history of southern England. While the length may be intimidating...

... examples with plasmids deposited at Addgene.  PRESTO-TANGO: The ligands that activate many GPCRs are...Roth’s lab created an open source resource called PRESTO-TANGO (parallel receptorome expression and screening...transcriptional output - TANGO) (Kroeze et al., 2015). PRESTO-TANGO uses a modified Tango beta-arrestin recruitment...ligands for unknown GPCRs. You can find the entire PRESTO-TANGO kit at Addgene! TRUPATH: Bryan Roth’s lab...McCorvy JD, Giguère PM, Sciaky N, Roth BL (2015) PRESTO-Tango as an open-source resource for interrogation...

...initial reversal drive cannot restore the exact original sequence, it can restore the original protein-coding...are eliminated by natural selection, gene drives distort inheritance in their favor and consequently can...they should not be relied on exclusively because historical studies of pathogen research have conclusively...using a recoding strategy; a subsequent drive can restore the wild-type sequence (save for the residual sgRNAs...

...Fast-FT, and mK-GO. Large Stokes Shift (LSS): Stokes shift (named after George G. Stokes) is the shift in wavelength... most FPs, Stokes shift is less than 50nm (often much less).  For LSS proteins, the Stokes shift is ≥ ...

... wrong. It brought the whole field to a grinding stop and only now, 2 decades later, is it finally back...showing that AAV-mediated delivery of the RPE65 gene restores vision in the dog model of LCA (Lancelot, one ... 10 more years to obtain FDA approval and make history in the form of Luxturna™. A better vector for ... again 3. Acland G. et al., 2001. Gene therapy restores vision in a canine model of childhood blindness...PubMed PMID: 11326284. 4. Bennett J, 2017. Taking Stock of Retinal Gene Therapy: Looking Back and Moving...

...when balancing a handful of demanding courses, a capstone paper I really wanted to hit out of the park, .... I need it just as much in my current role as Customer Support and Operations Manager at Addgene. In ...about to keep silent and move on to other things…. Stop! Take a step back and think of a time you’ve lead...collaborator or co-worker and showed that you really understood their side of an issue. Maybe you also showed...article from The Muse. Kai Strickland is a former Customer Support and Operations Manager at Addgene and ...

...Ampicillin Ampicillin, often just called amp, is the historical king of the antibiotic resistance realm. Amp ... factor. Zeocin is handy because it can be a one-stop shop for selection across systems and species. The...entirely detoxified, even when Sh ble is expressed (Trastoy et al). Thus, it may not be a good choice for experiments...Hadley, W. K., & Brooks, G. F. (1986). Effect of storage temperature and pH on the stability of antimicrobial...11, 56. https://doi.org/10.1186/1475-2859-11-56 Trastoy, M. O., Defais, M., & Larminat, F. (2005). Resistance...

...celebration, we thought it only fitting to share some stories of ways we’ve turned the available into the accessible...countries, import and export regulations, differing customs processes, and the distances the packages needed...other things, could ensure our packages navigated customs and arrived at their destination in a timely manner... to Design a Primer Creating Bacterial Glycerol Stocks Inoculating a Liquid Bacteria Culture Collections...scientists, from techs to grad students to professors, stopping by to say, “Everything I know about virus, I learned...

...chose needed to fulfill certain requirements: Customizable to our specific needs Modern and well-supported...the order and usage of the scripts needs to be customized in the pipeline for the specific needs of the...Since each process can use a unique container, we customized each container for its particular purpose and...bbduk process.    At first, to make sure we understood the concept of channels and implemented the input...user interface. In our next post, we’ll share our story about how we used Nextflow Tower to augment our ...

...yourself needing to brainstorm with a team. One major challenge with these brainstorming sessions is avoiding...with unlimited time? Another strategy for your brainstorm session is to start with something completely... ideas that I have found helpful in my team brainstorming sessions.  Allow the group time to think about...position or the longest tenure, but don’t let that stop you from speaking up.  If your ideas are not well...

...Stochastic multicolor labeling is a popular technique in neuroscience and developmental biology. This... persistently fluorescent during development. Stochastic multicolor labeling systems, many based on Brainbow...for improving the level of detail possible for stochastic multicolor labeling of the brain. Labeling Drosophila...express the same XFP transgene in an adult fly. As a stop cassette is located directly upstream of the fluorescent... researchers who developed Tetbow. Conclusion Stochastic multicolor cell labeling systems allow single...

...situ hybridization histochemistry: use for mRNA detection.” The Journal of Histochemistry and Cytochemistry...interaction has also been adapted for use in immunohistochemistry and antibody labelling. In these experiments...referred to as the Avidin-Biotin Complex (ABC) immunohistochemistry (IHC). For more information on ABC and IHC...Cytochemistry : Official Journal of the Histochemistry Society, 41.12 (1993): 1751–1754.PubMed PMID: 8245422. Additional...

...proteins from E. coli, Bacillus subtilis, or Pichia pastoris depending upon your specific protein folding needs...alleles in microbial populations through time, Christopher Marx's lab has engineered and validated a new...one-pot reaction. Using this system, you can create custom RNA binding proteins in as little as 3 days. Abil...Abil et al., 2014 show that you can fuse custom PUF proteins to a translational repressor to down-regulate...Rajarshi, et al. "Engineering RNA endonucleases with customized sequence specificities." Nature communications...

...papers or are brainstorming new technological innovations. Let’s take a look at three historical techniques...that save both time and money. Yet, knowledge of historical techniques comes in handy whether you are perusing...throughout the world. Let us know your favorite "Historical Lab Technique" in the comments section below!...Benjamin Chain. "The sequence of sequencers: The history of sequencing DNA." Genomics (2015). PubMed PMID...

...Science History Rap Battle A great example of science story telling following the unfortunate story of Rosalind...and their importance in understanding biological history. 4. Please Don't Kill My Hive Beautifully filmed...