...recombination scar encoding eight amino acids, but Golden Gate assembly can be designed to be scarless. Golden... in the final construct, allowing for precise, scarless cloning. The cloning scheme is as follows: the...

...ultimate goal to develop low cost and quick point-of-care diagnosis kits (reviewed in Tsang and LaManna, 2020...JC, Shon DJ, Fox DM, Puschnik AS, Bertozzi CR, Carette JE, Stanley SA, Harris E, Konermann S, Hsu PD (...Lelis FJN, Habibi S, Cai Y, Rennick LJ, Duprex WP, McCarthy KR, Lavine CL, Zuo T, Lin J, Zuiani A, Feldman...

...Conveniently, plasmids carrying genes for bacterial toxins like colicin typically also carry the necessary resistance...century! Nevertheless, the transfer of plasmids carrying these genes between bacteria can also have serious...

...advantage over Taqman is price point, which should be carefully balanced against the limitations of the assay....left, of droplet immersion technique, courtesy of CAR15.) Each droplet contains a primer and probe set for...Cloning Techniques References Bell AD, Usher CL, McCarroll SA. Analyzing Copy Number Variation with Droplet...

...which means it requires multiple Cas effectors to carry out genomic editing. Specifically, these types of... using a gRNA and can then recruit Cas3, which carries out the brunt of the mechanical work for editing...The reeling mechanism The reeling mechanism is carried out much like the name implies — after nicking,...

... Andrew and Maria were skeptical. The team was careful to test the possibility that this might just be...proteins that had shown magnetic responses: EGFP, mScarlet, and AsLOV2. After several rounds of semi-random...semi-random mutagenesis and screening, the EGFP and mScarlet variants were showing no obvious signs of improved...

...figure at right for an example). Look out for carbohydrates Certain strains, including HB101 and derivatives...derivatives, release large amounts of carbohydrates that can inhibit the cell lysis process. To ensure that..., buffering agents, and/or supplying additional carbon sources such as glycerol or glucose (in moderation...

...for viral genome sequencing (VGS). Why should you care about viral quality control? AAV vectors are produced... sequence. The resulting list of hits is then carefully reviewed. As mentioned earlier, it is common to...the contamination can be identified, and we will discard any AAV prep we suspect is contaminated. For AAV...

...Transformed cell lines Transformed cell lines Cargo Nucleic acids or protein Nucleic acids Nucleic acids...Microinjection Cells Many types Single cells Cargo Nucleic acids or protein Nucleic acids or protein... nondividing Dividing Dividing or nondividing Cargo Nucleic acids Nucleic acids Nucleic acids Throughput...

...spread even when costly. If even a single organism carrying a synthetic gene drive were to escape the laboratory...trust in scientists to develop new technologies with care and humility. Nothing would damage that trust more...aaa5945 (2015). doi:10.1126/science.aaa5945. PubMed. DiCarlo, J. E., Chavez, A., Dietz, S. L., Esvelt, K. M....

...high-throughput application is much more straightforward to carry out because it relies on a single BiFC pair. Find...Experiment Much like FRET, BiFC experiments must be carefully optimized to be successful. A BiFC experiment ...construction Fusing FP fragments to your PIPs requires careful selection of FP fragments and, preferably, some...

... settles in and people get the time recurrently carved out. This notice typically should include the paper...suited for different applications and should be carefully chosen for each group and topic. For example, ...purpose, journal clubs should be well planned and carefully organized. Most importantly, they should be regularly...

...micro, and cell biology are often more suited for cartoons or graphics for visuals. However, it’s much more...need text to properly contextualize. Graphics/cartoons (no words) Low Visually compelling and flexible...requires both artistic and scientific training. Cartoons may be read as “for children” or may need text...

...groups of trainees at an earlier point in their careers? A CURE for Biochemistry Undergraduate biochemistry... then proceed to use those skills to design and carry out their own novel experiments in semester two ...as they increase student interest in scientific career paths while also broadening access to experiences...

...bacterial CRISPR targeting. Phage-derived vectors carry CRISPR RNAs (crRNAs) targeting either genomic (blue... integrating into the bacterial genome, and it carries CRISPR machinery targeting 1) the given resistance...hardest-to-kill bacteria. In using phage-derived vectors to carry CRISPR, we’re turning the tables on the bacteria...

...generating interesting drug-like compounds especially cardenolides which have been used to treat heart disease....disease. Caterpillars that eat milkweed generate cardenolides that tastes yucky (and is poisonous) for bird ...system (Receptor-Mediated Ovary Transduction of Cargo)  delivers Cas9 RNP to the arthropod germline by...

...Cultures should only be passaged 20-30 times before discarding and thawing a fresh vial. In many labs, the history...Routinely test your cell line for mycoplasma and discard it if the culture has become overly confluent or...ensure high quality and reproducible results always care for your cell lines and virus properly and be consistent...

...’s which allow researchers to deliver oversized cargo (Hirsch et al., 2010). Today’s blog post will give... https://doi.org/10.1016/j.celrep.2017.06.064  Carvalho LS, Turunen HT, Wassmer SJ, Luna-Velez MV, Xiao...transduction following recombination between AAV vectors carrying parts of a larger gene. Nat Biotechnol 20:697–...

... a western blot — and why you might want to be careful relying on so called “housekeeping proteins.” Fully...approach is so common that we’ve used it for our cartoon gels so far. However, it throws both assumptions...Convenient! Comparison of loading control approaches Cartoons are nice and all, but we’ll leave you with this...

...in vitro translation system. The gene set was carefully chosen to perform statistically relevant benchmarking...conveniently performed through the N-terminal eGFP tag carried on all constructs. The Cell-Free Protein Expression... 48 plasmid adapter library, with each plasmid carrying a unique barcoded Illumina-M13F bridging primer...