...Addgene Blog Learn about Synthetic Biology with BioBuilder Use Addgene Videos to Learn about Basic Lab...genes in other species, but these studies generally leave questions about gene function wide open. What if...Check Out Our Molecular Biology Reference Pages Learn Simple Lab Techniques in Our Protocol Videos ...

...forming noncovalent dimers. This property of EGFP can lead to significant artifacts. If you're using green ...reportedly monomeric FPs are not actually monomeric, at least in practical terms. The propensity of a particular...FP dimerization coupled with POI dimerization can lead to polymerization of the fusion protein. A dimeric... members (superfolder, Emerald, CFP, YFP, BFP, Cerulean, Turquoise, Venus, and Citrine) can be monomerized...

...serotype, as some rAAV serotypes are abundantly released into the culture media, while others remain cell-associated...and non-genome-containing (empty) capsids, thus leading to an overestimate of transducible units in the...and dehydration (Howard & Harvey, 2017). You can learn more about common AAV applications by reading our...Wilson, J. M. (2010). Efficient serotype-dependent release of functional vector into the culture medium during...

...desired recombination event will remove NeoR only and leave exon 2 floxed, as seen in line 4 of Figure 4. Since...tools for creating knockout animals or cell lines, please consider sharing them by depositing with Addgene... with CRISPR through Non-Homologous End Joining Learn How the FLEx Switch Can Be Used to Turn Genes On...Mammalian Expression Vectors Find Cre-Lox Plasmids Learn More about CRISPR  ...

...remember what to say and provide your audience with a clear picture of your work. Practice, practice, practice...with people inside and outside your field. Your colleagues, lab mates, and PI can provide feedback on your...Anticipate questions: When practicing with your colleagues, ask them to brainstorm potential questions you... of a member of the Addgene depositor community Learn about careers at Addgene ...

...communication. But first, take a deep breath; at least you know you have an answer to that previous nosy... complicated process even more difficult. I’ve learned that some of the technologies that helped us navigate...application process and usually occurs in the few months leading up to applications opening. With virtual options...be a great opportunity to introduce yourself and learn more about your schools of interest. Benefits of...

...four times in a 95% ethanol and dry ice bath to release the virus particles from the cells. Cell lysates...batch-to-batch variability. Check out this post to learn more about different ways to titer AAV. Do you ...you have any AAV production tips or tricks? Please share them in the comments below! Special thanks ...first time CRISPR user Resources on Addgene.org Learn how Addgene produces AAV and lentivirus Check out...

... blog Learn the basics of Golden Gate assembly Check out NEB's work on ligase efficiency Learn about ...assembly, ligation of mismatched pairs of overhangs can lead to incorrect assemblies, so care must be taken to...is a molecular biologist and a Development Group Leader overseeing a team bringing new products to market...

...and controls (see Tips below), and by validating leads with other methods. More common are false negatives...levels of bait and prey proteins - overexpression can lead to false interactions. Lowered expression levels...techniques! Minimizing false negatives: Interactions leading to detectable reporter signals depend on a number...problems with one or more of these parameters can lead to a high number of false negatives in Y2H screens...

...a cooler. Chances are one of your colleagues has a frisbee at least. Use it! 8. Organize If you’re doing...make an effort to build relationships with their colleagues. It can be easy to get caught up in the minutia...true. If your event requires a lot of set up or cleanup, ask for volunteers to help out. Delegate- it will...

..., and they stepped up to apply for a grant which lead to their new career path.  A part of the career ...story for most established grant professionals is learning to succeed in grants through trial and error, ...increased their demand for stronger budgets with clear budget narratives, logic models, established partnerships... Throughout these moves and program changes, I learned to write grants for health, housing, economic development...

...research interests include proteomics, machine learning, and visualization for big biology. GPS for the...are critical? One hypothesis: the enhancers! This leads us to a three-step approach for designing an experiment...database. Aneesh's research interests span machine learning, proteomics, and user interfaces for math.     ...Catch Up on Your CRISPR Background with CRISPR 101 Learn about CRISPR Delivery Methods Validate Your Genome...

...numerous discoveries have been made using the Cas9 nuclease in the absence of genomic alterations. These studies...sequence via a targeting gRNA. But dCas9 does not cleave the DNA. Much of the research using dCas9 has focused...research conducted in the Hodaka Fujii Laboratory, please also see our interview with Dr. Fujii. References..., et al. "A programmable dual-RNA–guided DNA endonuclease in adaptive bacterial immunity." Science337.6096...

...innate immune system; and reconfiguring of the nuclear compartment, gene expression, and protein translation...responses that inactivate viral genomes or otherwise clear the virus from the cell population (Kafri et al....Adenovirus Tales: From the Cell Surface to the Nuclear Pore Complex. PLoS Pathog, 11(6), e1004821. https...stages of infection for efficient cell lysis and release of adenovirus from infected cells. J Virol, 70(...

...finding how-to videos on YouTube long before he learned (somewhat successfully) how to text or use Facebook... communication that you enjoy and that plays at least slightly to your strengths. Otherwise, you’re going...cost or freeware alternatives and how steep of a learning curve it will be to get to the necessary level...balance out cons. Flexible. Can allow people to learn in multiple ways from the same experience.  Can ...

...and ssDNA cleavage by canonical base excision repair enzymes. At the same time Cas9n cleaves the opposing...editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464–471 . https://doi.org/10.1038/...and enrichment of DNA editing by APOBEC–Cas9 or cleavage by Cas9 in living cells. Nucleic Acids Research...

...also reversible: once the environmental trigger leaves, gene expression is no longer activated. However...N2A, 3T3) and organisms (Chavez et al., 2016). To learn more about the different types of Cas9 activators....3149 Additional Resources on the Addgene Blog Learn how to Choose the Best Cas9 Variant for Your Next...More about Genome-wide Screens with CRISPR/Cas9 Learn How CRISPR Has Been Used with AAV Resources on ...

...elements, insulators, and silencers; however, some "leakiness" of transcription may occur. This is normally ...small RNA expression shRNA From the human U6 small nuclear promoter Constitutive  Murine U6 is also used, ...lacI or lacIq). Can be induced by IPTG or lactose. Leaky promoter with somewhat weak expression. lacIq mutation...the Addgene Blog Browse All Plasmids 101 Posts Learn about Mammalian Vectors Read about Reporter Gene...

... Make it clear that you are interested in the specific position you are applying to. Be clean and professional...letter, and if possible, send it to a mentor or colleague for a second look and helpful feedback!  What ...     Additional Resources on the Addgene Blog Learn to Develop Your Professional Network Become A Good...

... Flow data is messy, so learning how to gate takes a lot longer than learning how to read a gating strategy...visualization tool, a quadrant will often be too clean for real-world data. Most plots will have a shape...    Of course, there are many more things to learn about when diving into flow cytometry data. There...