...member of the Addgene Advisory Board, and Associate Director of the Genetic Perturbation Platform at the Broad...per gene. Further, the pools are available in two vector formats: all-in-one (lentiCRISPRv2, expressing ...both Cas9 and an individual gRNA from this single vector) and guide-only (lentiGuide expressing only an ...blogger, John Doench! John Doench is the Associate Director of the Genetic Perturbation Platform at the Broad...

...other researchers, and cloned them into lentiviral vectors to generate a kit which is now available at Addgene...cDNAs into a single consensus lentiviral expression vector and attached a DNA barcode to each to make the ...environments, for example. “We’ve been contacted by investigators all over the world who are interested to use...researchers have plans to expand the library to include activators for more cancer-related pathways and more nodes...

...yeast. These tagging vectors are based on previously described pFA6a-link vectors and include a Kan, SpHIS5...from these yeast-optimized fluorophore tagging vectors for your single or multi-color imaging experiments...protease site)-6xGly linker-(epitope tag)-ADH1 terminator-(selection marker). Additionally, Dr. Formosa...

... created the IBEX Knowledge-Base, a community repository and resource for spatial biology enthusiasts....includes 1049 community-validated reagents from 20 contributors, including 61 fluorescent probes, representing... showing the IBEX Knowledge-Base, with every contributor listed as an author.  The IBEX Knowledge-...Imaging Community’s vision is an open, global repository for reagents, protocols, panels, publications...

...have deposited their plasmids into the Addgene repository. More than 4,500 plasmids have already been deposited...aware of what Addgene can do for them and how the repository can help them share their materials. The new ...macrophages/monocytes using lentiviral expression vectors. I also used plasmids made and shared by others...carrying out my studies, and getting them from collaborators was extremely helpful, but with hindsight I ...

...pre-clinical and human trial phases.   So how does CRISPR factor into this workflow? CRISPR both speeds up the process...mutations and then test both individual and combinatorial drugs on for effectiveness prior to patient ...breast cancer and identified several candidate inhibitors of gene X in a screen. You generate a gene X ...CRISPR Guide CRISPR Plasmids and Resources CRISPR History and Development for Genome Engineering    ...

...bacteria, plants, and yeast. Popular cytosine base editor pCMV-BE3 has been requested nearly 400 times, and... plasmids covering three different epigenetic effectors. The diversity of tools is especially exciting... subpages, our updated CRISPR Guide and CRISPR History provide a great overview of the development of ...resources: CRISPR Plasmids and Resources and CRISPR History and Development for Genome Engineering  ...

...through Caitlin’s blood. “I come from a family of educators, mostly elementary school, middle school, and ...school, I actually served as the student course coordinator. With the guidance of one of the faculty members...their lecture, giving them advice, [and helped] proctor exams.” These teaching experiences led Caitlin ...interview experience.” Although luck wasn’t the only factor in Caitlin landing her job. All of the hard work...

... we’ve curated microbiology plasmids from the repository onto one handy Microbiology Resource page and...Addgene’s plasmid sharing services, as well as #happydepositors who stopped by to say hello. Needless to say...amazing work already represented in the Addgene repository (like these new Rainbow Bacterial Fluorescence...sphaeroides!) at the ASM booth. As well as being the Director of the Great Lakes Bioenergy Research Center and...

...How else can antibiotics be used in the lab? Historically, antibiotics have also been used to disrupt ...successful cloning. In these instances, the cloning vector typically has two separate resistance cassettes...powder form, but quickly breakdown in solution. Storing aliquots at -20oC and avoiding repeated freeze/...not add them to media hotter than about 55oC and store plates/stocks wrapped in foil if a light-sensitive...

...Addgene.org Find Plant Vectors at Addgene Find Simple Protocols for Introductory Lab Work MoClo Plants ... more than 100,000 samples to researchers and educators in 60 countries each year. Introducing Arabidopsis...Science and Industry (COSI) and the Franklin Park Conservatory. Do you use Arabidopsis in your research? Are...

...after subtracting out non-specific binding). Historically, glutathione-S-transferase (GST) fusion proteins...the extracellular domain of the erythropoietin receptor and subsequently evaluated interactions with mutant...antibodies against MBP are commercially available. Factor Xa loves calcium, but metal-chelate matrices, not...Addgene.org Learn about Popular Plasmids in the Repository Learn about New Plasmid Technologies in our Hot...

...the table below for more details.   Common Vectors Copy Number+ ori Incompatibility Group Control ...Relaxed This table defines common cloning vectors, their copy number, ori, and incompatibility groups... ori's are referred to as phagemids.   Other factors that affect plasmid copy number  Although the...affect the copy number of a plasmid, other external factors contribute as well. These considerations are especially...

...). Similar to the goodwill of Addgene plasmid depositors, this growing resource of over 100 entries is...data prepared. All of these questions relate to factors that you should have already recorded as good reproducible...Chemogenetics) or light (Optogenetics) to activate receptors? If you’re in the process of designing your experiment...Addgene.org Read our AAV guide Browse all  AAV vectors Find ready-to-use AAV preps ...

... able to eliminate the need for axenic (pure) laboratory cultures to study these microbes. From 16S rRNA...included a voltmeter on top of the battery box to monitor charge status and a disconnect switch between the... the battery bank and the vehicle battery/alternator to make sure the vehicle battery didn't run down.... can enable applications where reagents must be stored at elevated temperatures for long time periods ...

... first step. Developing mentoring relationships, especially with peer mentors, to discuss your strengths...recently, I have been hearing exit survey data from postdoctoral programs in the Boston area that demonstrate...Both explicit and implicit aspects of today’s postdoctoral training can directly interfere with a seamless...

... CRISPR system was involved. Today, that same laboratory enjoys a successful edit rate of over 90% in ...such as ApE or commercial workbenches such as MacVector or Benchling. Utility functions: These functions.... For this reason I generally recommend that laboratories that have already attempted several CRISPR experiments...above or, even better, first-hand knowledge from laboratories employing CRISPR on the same organism could ...CRISPRseek offers great utility and coupled with Bioconductor should be the first choice for anyone already...

...none are lost. Each Addgene library has its own depositor transformation and amplification protocol, but... a file in FASTQ format, a standard format for storing NGS reads and their corresponding sequencing quality...from the count_spacers.py file from Feng Zhang’s laboratory. There are a number of other analysis scripts...screening, and if you have any tips, input, or stories of interest about library use, let us know in the...

...Tools for checking grammar and jargon Hemingway Editor  This is a free web-based tool for detecting the... saved to your Mendeley library. The library is stored in the cloud, so you can access it from anywhere...you organize and cite papers. With the Zotero Connector, papers can be saved from Firefox or Chrome browsers...with others which makes it easier to work with collaborators. Zotero is a desktop based program that’s compatible...

... at 230 nm. The A260/A280 ratio is used as an indicator of DNA purity. Ideally, this number should be ...Concentration (ug/ml) = A260 reading x dilution factor x 50 ug/ml This method is quick and simple and ...like a PCR product). This method also gives an indicator of DNA or RNA contamination based on the presence...protocols and tips Find the best way to elute and store your plasmid DNA Learn about the different grades...