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...screens to identify genes involved in a phenotype of interest. This blog post will discuss differences between...target sites occur ~1 in every 8 nucleotides, restricting gRNAs to 5 – 65% of the protein coding region... the lab. Importantly, for any modification of interest, it would be unwise to make conclusions on the...experimental strategy is clear: for any gene of interest, one should require that multiple gRNAs of different...

... other molecule. When paired with a protein of interest, these tools provide an easier means to identify...as empty backbones to clone your own protein of interest. Fluorescence microscopy takes advantage of the...of cellular components, typically a protein of interest. Fluorescent dyes and ligands vary in their chemical...fluorescent ligands can elicit varying degrees of β-arrestin-2 recruitment to receptors, indicating their agonistic...

...comments. Additionally we’ve seen what topics are of interest to you, our readers. Below I’ve compiled a list... from the past year. A quick glance shows that interest in CRISPR continues to grow as the genome editing...community, so email us at [email protected] if you are interested. Addgene's Most Popular Posts of 2014 How...

...N-terminal tags, coding sequences of the gene of interest, C-terminal tags, plant terminators and resistance...component of Golden Gate and GreenGate are type IIS restriction endonucleases. They are like other endonucleases...their own lab: be very careful with the type IIS restriction endonuclease of choice, BsaI, which can be very...

...gene of interest in neurons. This ability to be paired with any Cre-controlled gene of interest makes pmSyn-BFP-Cre...may want to further control where your gene of interest is expressed.  A cell-type specific Cre can provide...

...two phosphatases, PTPN1 and PTPN2, which are of interest as cancer immunotherapy targets but are embryonic...s laboratory at Harvard Medical School and is interested in improving CD8+ T cell responses to cancer....generation of exhausted CD8+ T cell subpopulations and restrains tumor immunity. Nature Immunology 20, 1335–1347...

...expression. Cultures are then lysed and the protein of interest is purified using an affinity tag. Additional ...Determining the best way to express one’s protein of interest can save a lot of time later. Protein purification...C-terminal. Each set of plasmids uses the same restriction enzymes for cloning (see schematic below), enabling...

... to on Twitter? Prior to writing this post, my interest in Twitter was fleeting. I’ve had an account for...Unfortunately, it never clicked for me. Recently my interest was renewed in part due to FOMO  but mostly because... scientist? If you don’t use Twitter, are you interested in getting started? Reply in the comments below...

...With many interesting articles coming out in a myriad of journals every week, it can sometimes be difficult...plasmids and publications relevant to your research interests. We do this by promoting citation, supporting ...encourage you to fill out this form if you’re ever interested in writing your own blog post about a plasmid...

...to determine which job boards list positions interesting to you. Many job postings will result in hundreds...address how you are qualified and why you are interested specifically in the position. We frequently receive...cover letters. It is also important to express interest, even excitement, in a position. Everyone prefers...

...somewhat unusual things. For example, I am not that interested in meeting movie stars, but when I met the astronaut...a scientist up there, but the panelists were interesting enough to make up for the somewhat naïve questions...their seminal, cooperative paper in Science.  Interestingly, this project started with their labs studying...

...NEBCutter2: For finding restriction enzyme cut sites Webcutter 2.0: For finding restriction enzyme cut sites ... different types of cloning methods, such as restriction enzyme cloning, Gibson assembly, or In-Fusion...

... Short segment of DNA which contains several restriction sites allowing for the easy insertion of DNA....“vectors” or “constructs.” To insert a gene of interest into a vector, scientists may utilize one of a... a variety of cloning methods (restriction enzyme, ligation independent, Gateway, Gibson, etc). The cloning...

...figures of interest along with the article metadata, legend and user created notes on a “Pinterest for scientists... scientific literature? Very often the data of interest are in 1 or 2 figures from one paper and then ...

...2008 the Quake Lab at Stanford University became interested in exploring biological dark matter – large tracts...like YNP thermal sites - are also of practical interest since they’ve previously yielded valuable enzymes...bacterial RNA-Seq. This enriches for transcripts of interest without an increase in the number of steps in ...

...collection, you'll likely find some proteins of interest for which you have several antibodies in stock...another, or if you’re looking for a protein of interest while it’s in a transition state or specific configuration... to validate that it recognizes the protein of interest in specific configurations or conditions. In those...

...easiest and clearest ways to determine if your sample has recombined. Choose a restriction enzyme that...one resulting plasmid carries the transgene of interest (aka the viral genome) and the other contains ...

...overcome. Conflict, and its inherent tension, is interesting. It’s what drives a story forward - and this ... the pieces that make you think, “Oh, that’s interesting! How did you solve that problem?” Those are the...your audience in with something that sparks their interest and makes them want to know more. This could be...

...plasmid that produces biotin ligase and a protein of interest fused to maltose binding protein (MBP, New England...-Avi-His/BirA to add biotin to your protein of interest Grow fusion protein plasmid in NEB T7 SHuffle... Vanderbilt Eye Institute and is particularly interested in cell signaling.     Additional Resources on...

...several excellent guest posts from our depositors. Interested in writing your own post for the Addgene Blog...(AAVs) are commonly used to deliver a genes of interest to a variety of cell types in-vivo. These are ...customizable functions to your DNA sequence of interest. Potential applications of the system include ...