... here!). The only issue with these GEVIs? They perform poorly in two-photon conditions, which are necessary...the fluor via FRET. This system is particularly powerful for in vivo imaging especially since JFs can cross...type of interest. Viral pseudotyping is also a powerful tool that swaps capsid proteins from different...

...monomeric FPs. EGFP and other GFP family members (superfolder, Emerald, CFP, YFP, BFP, Cerulean, Turquoise,...dimerization. Importantly, all of these assays are performed in vitro. Assays to detect FP dimerization in ... 1 and 2 depend on protein concentrations. An imperfect solution is to restrict imaging analysis to cells...

...support your professional development I had a wonderfully supportive mentor and graduate office. I cannot...ages, learned a lot about the dinosaurs, and met wonderful people. When I started volunteering, I was worried...Scientists Read Emma Markham's Post on Finding Your Perfect Job After University Browse All of Our Career Posts...

...One of the most powerful strategies to investigate a gene's function is to inactivate, or "knockout",...arms. When we select cells later, we will first perform a positive selection for the antibiotic resistance... knockout and new CRISPR tools continue to be perfected, the production of cells and mouse knockout lines...

... have a much better chance of creating a high-performing team - which is every manager’s dream. Learn ...often associate feedback with end-of-the-year performance review, but feedback should be given throughout...the year and in a timely manner. If someone’s performance or behavior is inadequate, you need to address...

...the genome. Let’s now examine how we can locate powerful stretches of DNA known as enhancers with the help... the techniques from Table 1, provides us with powerful GPS-like techniques to explore dark matter in ...interests span machine learning, proteomics, and user interfaces for math.     Additional Resources Beyond the...

...contamination could interfere with interpreting the results of transfection assays. Go forth and perform your DNA...when the kits buffers ran out. Homemade buffers performed the same as those provided with the kit. Although...

...first note that quantitative PCR (qPCR) has been a powerful tool for quantifying AAV for some time now. When...readout of your samples.  Of course, if qPCR were perfect there would be no need for ddPCR. One issue with...Furuta-Hanawa et al., 2019). Tips and tricks for performing ddPCR Clean everything You must have a clean ...

...Addgene’s repository) and make it easy to find your perfect match. Choosing a Cas9 for your knockout experiment...deaminase to where the edit is desired, and it performs the programmed edit within the editing window ...with this tool is that the desired edit will be performed on all relevant bases within the editing window...

...Wild-type AAV2 has been reported to have a near-perfect physical-to-infectious particle ratio of 1:1 (Zeltner...requires an electron microscope, it is not routinely performed on all new viral vector preparations. Addgene ...PMCID: PMC3706409. 7. Zeltner, Nadja, et al. "Near-perfect infectivity of wild-type AAV as benchmark for infectivity...

...Writing a review article is a wonderful way to develop and exercise your scientist skill set. If you ...you're becoming an expert in your field and are the perfect person to be writing the review! Doing so is a ...life’s work: so remember that done is better than perfect. Good luck! References 1. Haery, Leila, Ryan C...

...cannot be ignored. They have the potential to interfere with the function of resident ER chaperones and...and retained in the ER. FPs have been undeniably powerful and successful tools that have enabled numerous..., Lindsey M. Costantini, and Erik L. Snapp. "Superfolder GFP is fluorescent in oxidizing environments ...

... to the consistency of the reagents and hands performing the experiment, not the reproducibility or consistency...independent aliquots of cells and plasmids, be performed on different days, etc, but outside constraints...incorporating additional positive and negative controls, performing multiple replicates, and ultimately getting interpretable...

... may have contributed to this mouse-specific performance, why LY6A is an unlikely receptor for PHP.B, ...selection of a virus with mouse-strain dependent performance. This surreptitious finding illustrates how the...2019): 538421. If you’re interested in how PHP.B performed in non-human primates, check out these two references...

...analysis is performed yielding a graph of edits in the input population or clone.   How to perform a TIDE...homology-directed repair events, or maybe you’re performing a single base pair edit with a prime editor. ...

... GFP is such a well-validated, all-round good performing probe, it is the first choice when selecting ...reduces its size. The substantial size of GFP may interfere with the activity or the localization of the fusion...C-terminus of another protein. This may, however, interfere with the function of the protein. This is clearly...

... virus directly using saliva without having to perform RNA extraction but that may work only if there ... Motif) for target cleavage. This test can be performed in two steps for challenging samples as in Two-Step-SHERLOCK...fluorescently labelled substrate, gets activated and performs non-specific collateral cleavage. This cleavage...

...When performing restriction digests on plasmids at Addgene, we sometimes observe something odd in our...Running undigested DNA on a gel as a control when performing a diagnostic digest is a simple but effective... sequencing Addgene uses Illumina MiSeq NGS to perform whole plasmid sequencing of our plasmid samples...

...can be combined with CRISPR/Cas9 to efficiently perform genetic manipulations in animal models or cells...activity. In summary, the piggyBac™ transposon is a powerful technology that may be used for a variety of research...piggyBac™ for gene editing allows you to efficiently perform seamless genetic manipulations, while avoiding ...

...Read on for an in-depth breakdown of how to do perform restriction digests.   Overview of the restriction... common chemically-competent cells. You should perform, at minimum, two transformations after a ligation... 101 Posts Resources on Addgene.org Find the Perfect Empty Vector for Your Cloning Experiments Browse...