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...these subpages, our updated CRISPR Guide and CRISPR History provide a great overview of the development...characteristic probably explains why pC016 - LwCas13a guide expression backbone with U6 promoter has already...member Alex Chavez) Day 11: Pooled libraries: Genome-Wide Screening Using CRISPR/Cas9 and Plasmids 101: NGS...

...This is where affinity tags come in – they are peptides which do not naturally exist in most expression...antibody is an alternative to amylose resin and also provides a visual detection mechanism via applications ...clean up step of the tag – a handy feature. If you decide not to cleave the GST tag after/during purification...Addgene.org Addgene antibodies Addgene’s antibody guide Antibody protocols  Resources on the Addgene blog...

...short enough that everyone stays on the same page. Ideally, these meetings include all collaborators, from...meeting has an equal opportunity to contribute. Using video conferencing tools like Zoom or Google Hangouts ...pace at which problems can be addressed and new ideas generated. Furthermore, this casual communication...and grant applications in real-time so they can provide input at the times that work best for them. This...

...themselves. Therefore, your cloning strategy needs to identify cells containing the plasmid construct you’re ...positive selection strategy, here are a few to consider. Antibiotic selection Selection is straightforward...backbone contains your gene of interest. Auxotrophy Besides antibiotic selection, you can select for a successful...biology reference Watch molecular biology protocol videos ...

...software. Vectr has layers, which let you lock and hide individual vectors (shapes defined by 2D points ...OS X, and Linux. Inkscape is versatile and has a wide selection of tools for drawing and editing shapes...which requires users to give appropriate credit, provide a link to the license, and indicate if changes ... for reuse with modification. It’s always a good idea to double-check the image’s license information ...

...biggest advantages of such a tool is being able to identify specific transducer complexes that are activated...was more challenging. The Roth lab used structure-guided protein engineering and experimental refinement...protein signal transducers. The TRUPATH collection provides scientists with a toolkit to gain an understanding...Kadowaki S, Haga T, Takaesu H, Mitaku S (2002) Identification of G protein-coupled receptor genes from the...

...deposited at Addgene enhance scientific sharing worldwide but a select few are particularly special. Over...molecular tools with the scientific community worldwide, making it easier, cost effective and faster; ... Currently, there are roughbly 400 depositors worldwide who have one or more blue flame plasmids, with...literally enabled sharing with 100's of investigators worldwide. Now everything from every paper is routinely ...

...Molecular cloning techniques are some of the most widely used techniques in the biological sciences and ...unwanted mutations due to excessive PCR cycles or low fidelity Taq DNA polymerase, the construction of specific...during PCR or any subsequent steps. Using high fidelity DNA polymerases, I have never had any mutations...Amplification efficiency is highest when using high fidelity DNA polymerases like phusion. An 18-cycle PCR ...

...Career: Doer, Manager, Leader” by Charlotte Beers, divides workers into three cycles: Doers, Managers and ...motivate their teams, it is the leader’s job to provide the driving inspiration, to create a productive... Understanding these three roles allows you to identify what you need to do to be successful in a specific...bringing and effecting solutions to both known and unidentified issues. There are many good resources on the...

...their work.  Check out Joanne's Reddit AMA   Guidelines for giving feedback Use formal and informal ...real change is possible in your eyes so their confidence is restored. A good manager does not hold grudges...best? What 3 things worked least well? Manager provides feedback (may be the same as self-assessment or...about real performance issues. Watch this little video to learn more about indirect language from Steven...

...attention and enjoy anecdotal forms of information — identifying with a character, building emotional investment... do and should tell stories all the time. They provide accounts of their research struggles in papers.... a weekly lab meeting as you’ll need to for an outsider. And you won’t need to dive as deep into your ...preprint servers, share your email address far and wide, keep your website up to date, invite people to ...

...explaining a technical piece of research to others outside that specialty, who may be other scientists, or...journalists, or policy makers. The term editor covers a wide variety of careers and jobs. At Science, which is...financially critical. Those who work on the publishing side of science have professional meetings, societies...in the science communications world, you might consider an internship or a short program to build skills...

...Taylor, 2014). They’re so simple that we can’t even decide if they’re alive or not. Yet these simple, small...Virus structure While all viruses have a genome inside a protein shell known as a capsid, some also have...march to the beat of their own drum. They have a wide variety of possible genome types that can be either...copies, which are released from the cell like baby spiders from an egg sac. (Okay, some enveloped viruses ...

... the literature and find an ideal antibody against Protein A and an ideal antibody against Protein B. ...specificity to the target while constant regions are identical for antibodies within certain classification groups...with human antibody sequences.  Additional considerations Although isotype conversion is routinely used...

...know all about antibiotic resistance genes, let’s consider another basic element of any plasmid: the origin... intend to use, and whether or not you need to consider your plasmid's compatibility with one or more ...copy number varies. See below for additional considerations. *Requires pir gene for replication (reference... external factors contribute as well. These considerations are especially useful to keep in mind if you...

...Addgene closed for almost two weeks and with labs worldwide closing for unknown amounts of time, we were facing... enhances productive collaborations as well as provides visibility for grant-driven research being done...each of many grantors supporting research could provide Addgene with a small amount of funds, the community...and projects to serve the needs of scientists worldwide with Addgene’s traditional excellence through ...

...what gets packaged into the AAV molecule. What’s outside of the ITRs does not get packaged in the AAV vector...the WPRE. TL;DR? Watch our AAV Transfer Plasmid video!   Now you’ve got a basic understanding of some...Lisowski L, Alexander IE, Logan GJ (2019) A User’s Guide to the Inverted Terminal Repeats of Adeno-Associated...basic viral vector elements Read about four considerations for the perfect AAV tool Learn about synthetic...

...hybridomas) that secrete antibodies in culture and provide renewable reagents with reliable and reproducible...first NIH grant that Addgene has received. Addgene prides itself on being financially self-sustaining through..., Addgene’s interim Executive Director. “NABOR provides a new model for the creation and acquisition of... enable researchers everywhere.” The timing is ideal as the NIH’s BRAIN Initiative has been funding laboratories...

...quick look at your order status on addgene.org identifies the hold-up. Your Material Transfer Agreement...quickly! If your TTO receives a Golden Stopwatch, consider sending an email to thank them--after all, they...unprocessed agreements slow down your research; provide specific examples if possible Tell them about the...exchange of intellectual property (as inventions, ideas, plasmids) among universities and other research...

...lysosome, and degradation of autophagic substrates inside the lysosome. Historically, methods for studying...dynamic process into a static measurement and it provides limited information about what materials or organelles...autophagic activity. The aim of this post is to provide an overview of four autophagy biosensors currently...protein plasmids and resources Fluorescent protein guide: Biosensors ...