...neuronal networks Double inverted vectors expressing light inducible ion channels (plasmid # 20298 for example...manipulations in vivo, FLEx vectors expressing the light activatable ion channel under the control of Cre...proteins (e.g. postsynaptic PSD95-mcherry) are used in high concentrations in combination with low concentrations...vectors expressing the required recombinase. The high concentration of FLEx vectors guarantees that most...

...optimized for each specific virus in order to generate high quality preparations. After production, all virus...obtained using ddPCR are generally two to three fold higher than those achieved using the standard qPCR method...performed by ddPCR.  Quality Control Addgene ensures high quality viral vectors by optimizing and standardizing...staining or SYPRO Ruby staining and the molecular weight and relative intensity of the viral capsid proteins...

...choosing a quality mentor, and rightfully so! They can provide you with high-level career advice, open doors...won’t be very proficient afterwards. Below we highlight how to prepare and things to keep an eye out for...

...where the brain fails to divide properly into the right and left hemisphere. Addgene has several plasmids...Spaghetti squash (Sqh) is the Drosophila Regulatory Light Chain (RMLC) of nonmuscle myosin II (Karess et al...PMC2395242. Jordan, Pascale, and Roger Karess. "Myosin light chain–activating phosphorylation sites are required...12490562. Karess, Roger E., et al. "The regulatory light chain of nonmuscle myosin is encoded by spaghetti-squash...

...times during a 10 minute conversation. Leaving a tight-knit team like that can and will be a little gut-wrenching...camaraderie again in a non-academic career. You might not be in a lab anymore, but you will very likely...academia provide a formal training regime and the right set of tools to help you do your job as well as ...lab whether you realize it or not. These skills might include collaborating, managing, and communicating...

... intensity was classified into four groups (4; brighter than control, 3; same as control (Cetn1/sgRNA1...sgRNA1), 2; darker than control, 1; very dark). Brighter fluorescence indicates a more successful gRNA design...your cells under a fluorescence microscope. The higher the amount of fluorescence, the better that gRNA...been inserted or deleted. Each lab will use a slightly different method to genotype their pups. Taking...

... viruses to study. Interestingly, due to certain high profile problems surrounding cell culture over the...but these days it’s never been easier to conduct high quality work in cell culture. The problems with ... Fortunately it has never been easier to conduct high quality cell culture. Biorepositories around the...AstraZeneca-MedImmune, and Infors & Applikon. Due to his high level of experience he now travels around Europe...

...overview of plant genome engineering using CRISPR, highlight the specific modifications to CRISPR machinery...Lab.  As with all plasmids in the repository, we highly recommend reading the associated publications or...10.1104/pp.15.00636 Ma X, Zhang X, Liu H, Li Z (2020) Highly efficient DNA-free plant genome editing using virally...

... issue of “tiny vector, big transgene” by just straight up packaging an oversized genome of ~9 kb into...Generally an overlapping split vector has equal or higher transgene expression when compared to using a single...of non-endogenous sequences, such as a 77bp long highly recombinogenic homology sequence derived from the...method While the hybrid dual vector system has a slight advantage over the overlapping and trans-splicing...

... can find his original blog post below! --Ed.  Eight years ago, I decided to write a "how to" manual ... and mentoring to several people that found my insights helpful. All went on to get faculty positions ...potential colleagues study. Think about how you might collaborate or, at the absolute minimum, can appreciate...free!) contains strategies, hints, anecdotes, and insights. After I wrote the first version of my manual,...

...developed, which differ in the number, stability, and brightness of the fluorescent proteins used in the system...Brainbow 3.0 (Cai et al., 2013), incorporates the highly photostable fluorophores mOrange2, EGFP, and mKate2...system to boost XFP expression. The increased brightness of Tetbow allows this system to be used to trace...Sakaguchi, Richi, Marcus N. Leiwe, and Takeshi Imai. "Bright multicolor labeling of neuronal circuits with fluorescent...

...Biotin protein ligase (BPL) is responsible for this highly specific post-translational protein modification...photoreactive arylazide group. Under strong visible light, the arylazide group becomes reactive and forms ...: 20353954. 5. Kay, B., Thai, S., & Volgina, V. “High-Throughput Biotinylation of Proteins.” Methods Mol...PMID: 2388593 7. Holden, M. A., & Cremer, P. S. “Light activated patterning of dye-labeled molecules on...

...news briefing, to quick notes, to finances, to flights & ordering food - everything is covered.   So why...for a large institution or company, site licenses might be available as well. In most cases this applies...are collaborating with other labs or clients, you might need share data with them including reports and ...does the ELN provider protect and respect your IP rights and data ownership? IP and data ownership should...

...donor, you can deliver a silent mutation in a neighboring nucleotide along with your desired edit. This...provided you have some idea of where in the genome you might expect off target editing to occur. Luckily, in ...you to Sanger sequence a manageable number of the higher probability locations. If you’re worried about ...Bombieri N, Giugno R, Pinello L. CRISPRitz: rapid, high-throughput and variant-aware in silico off-target...

...to make them serve you well. The happy byproduct might just be a career opportunity. The focus of this...content. Create a Google.slides or Powerpoint file right now to capture interesting key results, striking...need to define a term, but are worried that this might sound condescending, consider explaining the term...Additional Resources Joanne's "How to Choose the Right Next Lab or Workplace" On YouTube Joanne's 10 Things...

... from a default OFF to an ON state, and how you might use these in your research. Today, we’ll look at...Take a look at all of the options and decide what might work best for your experiments! This post was originally...References and Resources References Gossen M, Bujard H. Tight control of gene expression in mammalian cells by...PMCID: PMC49329. Triezenberg SJ, Kingsbury RC, McKnight SL. Functional dissection of VP16, the trans-activator...

... site.  T-RL= transposon right-left orientation. T-LR= transposons left-right orientation. Image from ...2009). hySB100x improves on SB100X by having 30% higher transposition activity (Voigt et al., 2016). Sleeping...sequence into the zygote’s genome, a process that’s highly unpredictable. Transposons, however, efficiently...

...backwards. To publish a submitted manuscript in a high-profile journal typically takes 6-12 months depending...technology may not exist to answer the question or it might be extremely difficult for you to get the needed...for a summer undergrad, rotating grad student or high school student. Learn to teach how to design, perform...improves your perspective like intense exercise. I highly recommend running or biking or something that demands...

...activated, the protein emits a brief yet bright burst of light, visible to the naked eye, at regular intervals...forever change the fly lab experience. The Frequency Light Interval System, or FLIS, incorporates a bioluminescence...

...Addgene we couldn’t be more excited! As a small, tight-knit organization we thrive on 3 things: fun, teamwork...trophy among 6 people, but you’ve earned bragging rights for the next 12 months. Your team’s creativity,... favorite lunch... a BonMe noodle meal, crammed right into the box. Which is your favorite team costume...