...those used for breeding heterozygous mice. In this case, your parental mouse should be hemizygous at the...similar epigenetic modifications. Special breeding cases Whether your mouse colony consists of genetically...explained more in Part 1 of this series). In any case, Cre is a dominant phenotype, while you’ll want ...

...your gene of interest.  SapTrap Tool Kit for CRISPR Cas9 edits At Addgene, we have the entire SapTrap toolkit.... "SapTrap, a toolkit for high-throughput CRISPR/Cas9 gene modification in Caenorhabditis elegans." Genetics...blog Learn more about SapTrap Read about CRISPR/Cas9 in C. elegans Browse our Plasmids 101 blog posts...

...single 72h harvest is best for larger genes such as Cas9. See figure 1 for an example of how changing transfection...enough to get a sufficient titer; this is often the case with large genes. This is due to inefficient packaging...inserts; a study by Kumar et al. found that in some cases, titers can drop 10-fold for every 2kb increase ...

...other things, science communication comprises broadcasting, science writing, and certain aspects of art...audio or visual person. Why don’t you record a podcast or put some effort into making a video? Use your...Communication Posts Browse our Career Page Listen to Our Podcast Episode on Science Communication Additional Resources...

...they’re knowingly selling a bad product (in which case, you should probably stop using that company). Frequently...frequently asked by customers (and I expect this is the case at most companies). Always go back to the source...that could take weeks to resolve adequately. Occasionally there are some really off-the-wall questions...

... matches the copy number of your sample. In that case, an RQ value of one (or, more realistically, between...ddPCR may be a better option for more challenging cases.  Price point If budget is a concern, look for...number and it may not be able to identify trends in cases where the copy number variation is two or less (...

...endoplasmic reticulum localization, Munro and Pelham casually introduced the Myc-tag (Munro, et al. 1986; Munro...developed almost 40 years ago? Well…kinda. In some cases, antibody providers are distributing monoclonal ...same hybridoma developed by Evans, et al. In other cases, providers have moved to producing anti-Myc [9E10...

...always maintain a backup replicates of each clone in case of contamination. We do not normally use antibiotics...well plate for further analysis. To assess CRISPR/Cas9 knockout efficacy, we have used both Western blotting...potentially large number of single clones obtained (on occasion we have prepared two 48-well plates of clones)...

...individual expression cassettes, having the genes expressed from the same cassette is sometimes advantageous...and you can then insert the whole multicistronic cassette into a backbone as a single unit. Although it ...

...compared the number of plasmid orders and new COVID-19 cases for each week by location. We observed that as expected...drop dramatically when there are many new COVID-19 cases being reported (right, red), likely due to quarantining...map showing the plasmid orders and new COVID-19 cases each week. Statistical Modeling Note -- If statistical...

...options available to today's science communications: podcasts, blogging, videos, lectures, exhibits, social ...content.  What are you interested in? Math, sarcasm, romance, and spacecraft! Image courtesy of xkcd.com...can increase exponentially with complexity. Podcasts Medium Easy for audience to engage while multitasking...

...zinc-finger nucleases, TALENs, and more recently CRISPR/Cas9. Another popular targeted gene deactivation method..., relying on established reagents such as CRISPR/Cas9, Tol2, and the high-quality sequenced zebrafish ..., and their resistance (so far) to facile CRISPR/Cas9-mediated insertion of foreign DNA. Despite these...

...applications, such as protein isolation from serum. In this case, Protein A and Protein G will bind both the primary...indiscriminately, causing competition for binding sites. In cases like these, antibodies can be directly conjugated...simply isn’t available for a specific target. In cases such as this, an epitope such as c-Myc, GFP or V5...

...Alongside there is a second-line team that takes on cases requiring in-depth analysis, data review, in-house...knowledge and judgement. For particularly tricky cases, team discussions can often lead to otherwise unobtainable...patient, attentive, empathetic, and tactful. Occasionally it can also be necessary to take the lead and...

...approach known as multimodal therapy. Sadly, in many cases the cancer is diagnosed so late that these treatments...success in treating mesothelioma, such as in the case of Paul Zygielbaum. A different approach to immunotherapy...and used throughout the world. New mesothelioma cases could be greatly decreased through a ban on asbestos...

...their data months or years before others. In some cases, typically consortia involving genomic screens, ...the originating researchers, even if they are occasionally beaten to press, as there is so much analysis...track the motion of multiple animals on different occasions via video, generating huge amounts of data. When...

...complement each other to restore the full-expression cassette. AAV’s natural ability to reassemble fragmented...sequential homology directed repairs (HDR) with CRISPR/Cas9. Donor A contains: 400bp homology arms to the site...2017) CRISPR-Mediated Integration of Large Gene Cassettes Using AAV Donor Vectors. Cell Reports 20:750–756...

... an intact sample, a cell in the case of ICC or a tissue in the case of IHC. IF is used specifically when...interest within an intact cell, tissue, or, in some cases, a whole organism. But, as with any antibody-based...

...(Mouse IgG2a) Goat anti-mouse IgG2a CASPR anti-CASPR mAb K65/35 (Mouse IgG1) Goat anti-mouse...Neocortex labeled with anti-pan-Nav (magenta), anti-CASPR (green), and anti-Kv2.1 (cyan). (B) Cerebellum labeled...

...plasmid with repeating sequences. In some specific cases, multimerization has been observed to occur more...reduce the frequency of multimerization, and in some cases completely prevent the formation of plasmid multimers...Sharratt, 1984; Summers et al., 1993). In these cases, screening multiple bacterial colonies using one...