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... oxygen concentration may affect photobleaching rates (Shaner et al, 2008). This brings us to the key ...application (Goedhart et al, 2012). The third issue relates to the environment of the fluorescent protein. ...The issues with quantification of photobleaching rates can be summarized as follows: Instead of determining...

... cross-talk with the other orthologs. Ma et al. tested their dCas9 variants using gRNAs specific for telomeric...individual colors to 6. A gRNA with all three hairpins generates white light. All 6 gRNAs can be expressed in a...separate vector. Figure 2: CRISPRainbow generates 6 unique colors and a white composite. gRNAs with...

...a new legal minimum of $47,476 per year, under updates to the Fair Labor Standards Act (FLSA). This was...22nd a preliminary injunction against the FLSA updates was granted nationwide. Again, we at Future of ...getting paid, which we will be analyzing for future updates. But suffice it to say, salaries are showing surprisingly...

...CASANOVA responds to blue light quickly. Within minutes of inactivating the CASANOVA complex, fluorescent...genes-of-interest (GOI) into two independent expression cassettes on a single plasmid (Grefen and Blatt, 2012). ...proteins that bind to either the light- and dark- states of the photoswitchable cPYP, which changes in conformation...

...caveolin 1, also known as CAV 1. Usually, caveolin 1 promotes the cell life cycle progression and enhances tumor... TAK 1, an enzyme that controls apoptosis and regulates transcription. Without TKA1, cells proliferate... 2004). In fact, Sanford H. Barsky and his team tested the influence myoepithelial cells had on the conversion...

...keep track of how you met the connection, as these notes are saved in the messaging stream. Unfortunately...LinkedIn is not the platform for politics, family updates, or click bait of any kind. For more on inappropriate...However, do comment on other people’s news and updates. Share your own good ideas, events and favorite...

...antibiotic in your plate might not work. Check the plates! Streak on them an E. coli  strain known to be ...growing fine, it is probably time to prepare new plates with a fresh stock of antibiotic. Many small colonies... different temperature (30°C) for the growth on plates and in liquid culture. If you don’t get any positive...

...and disease states, their circadian regulation, as well as in important physiological states such as adipogenesis...therapeutic indices, including hypertension, cancer, diabetes, cardiovascular disease, cholesterol gallstone...

...incompetent by using four plasmids instead of three and eliminates the requirement for Tat. Always includes deletion... transfer plasmids may have limited restriction sites or may only be compatible with certain cloning methods...target species. Although chromatin availability facilitates integration, it does not explain the lentiviral...more accurately predict and understand integration sites have led to the development of multiple genome-wide...al. (2020) for examples). Pseudotyping Tropism dictates which types of host cells the lentiviral vector...glycoprotein — transduce airway epithelium and cardiomyocytes in utero Rabies glycoprotein — transduce neural...for genome-wide mapping of lentiviral integration sites. Molecular Therapy — Methods & Clinical Development...

...animals may be variable among animals and bleed dates and even clonal hybridoma cell lines can produce...Tran et al., 2016). Each antibody would then be tested for antigen binding. If starting from an established...data (Begley & Ellis, 2012; Baker, 2015). This contributes to an enormous amount of wasted money and time...

... plate format yielding anywhere from two to six plates of 96 samples per week. This is where our new process... process only takes about one day, even for six plates (576 individual plasmids)! These libraries are ...takes about two days to complete. After the run completes, we begin our assembly process. Again, thanks ...

... used to integrate the tagged AID* into genomic sites of interests via CRISPR techniques. This tool-kit...activated, Cas9 cleavage can induce DSBs within minutes. New CRISPR plasmids for Borrelia burgdorferi from...are now available. These libraries encode target sites flanked by randomized nucleotides in place of a ...

... experiments is cutting efficiency so that the greatest effect possible will be achieved. On the other... an NG PAM.   Cas PAM sequence  Additional notes SpCas9 NGG - Cas12a TTTV Great for AT-rich...coverage in the human genome. SpRY Cas9 has PAM sites covering the majority of the genome! Want to...

...co-incubate it with an “assist peptide” that facilitates endosomal escape. Combining Cas9-PAGE with lentiviral... in rodents have not been widely successful in primates. The authors used a multispecies approach to generate...endothelial-specific tropism in rodents and broad tropism in primates. Nature Communications, 14(1), 3345. https://doi.org...

... high complexity Golden Gate Assemblies. Image courtesy of New England Biolabs.   DAD Tools The three...input a sequence, SplitSet finds optimal fusion sites to split it into multiple fragments. You can define...sequences or set a narrow search window for a site—or sites—that must be used. Each of the tools works with ...

... real challenge is finding relevant papers. The latest installment of the Early Career Researcher Toolbox...around since 1996, but is getting an update! The latest version can be found here. The new PubMed will ...cite the most), and who the author influences (who cites the author the most). Best uses Finding papers frequently...

... Viruses are intracellular parasites and natural vehicles for genetic information. Therefore they make...Addgene: Follow our viral vector blog posts for updates on the most recent deposits, protocols, and tips...MuLE (Multiple Lentiviral Expression) system facilitates the simultaneous introduction of multiple genetic...

...BioID, BirA* is fused to a bait protein and it biotinylates proteins that are within a ~10 nm radius of ...the bait proteins were tagged with. It’s best to test all combinations and pick the conditions that give...interactome. Further characterization of these candidates is critical to understanding how they interact...

...identified and interpreted in less than a couple of minutes. How does one accomplish all of that? What follows...The UCLA Library offers some Powerpoint based templates with different column numbers for a good starting...starting point. Adobe also offers free poster templates, although most are vertical.  Large central box...

... based off of p24 quantification tend to be overestimates since they can include free p24 and defective...vectors carrying fluorescent markers are used as it eliminates the need for antibody staining. Check Out Our...multiplicity of infection (MOI) to ensure only one copy integrates, selecting by antibiotic-resistance or single...