...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...

...thousands of designs in parallel. To do this, we performed high-throughput ‘self-targeting’ screens with ...PRIDICT2.0 score. For the best prime editing performance, we suggest that users select some of the best...learning–based model with high generalization performance. Science Advances, 5(11), eaax9249. https://doi.org...

... 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...

... capture of chromatin interactions The Xu lab performed a proof-of-concept experiment to show that CAPTURE...original CAPTURE method. Both CAPTURE methods outperformed other 3D chromatin methods such as ChIA-PET ...Wang, K. (2018). 3C and 3C-based techniques: the powerful tools for spatial genome organization deciphering...

...consider Gibson cloning. In this technique, you first perform a round of PCR to generate DNA fragments with 15...function while 5’ adenosine overhangs left by Taq perfectly anneal with the topoisomerase I generated overhangs...available cloning kits that one could choose from to perform these experiments. In most cases, there will be...

...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...

...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...

...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...

...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 ...

...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...

... 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...

...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...

...methods can interfere with binding, and some common antibody buffer components can interfere with conjugates... non-overlapping epitopes so that they do not interfere with each other. The detection antibody may either...

...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...

...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...

... (am): Observe fluorescence, harvest cells, or perform your experiment Equipment Class II, Type A2 Biological...

...relatively simple procedure and is useful for performing low efficiency transformations. Last Update: ...

...mummification). Permeabilization, as the name suggests, perforates the cell membrane to allow for antibodies to ...day. Note that intracellular labeling must be performed only after viability staining and surface labeling...surface labeling. Intracellular labeling can be performed on cytosolic and/or nuclear markers. Lastly, stains...