...easiest way to make a knock-in cell line is to utilize the built-in repair pathway cells already have ... multiple rounds of cleavage at the target site until either the PAM site or part of the guide’s recognition...the most common editing pathway for DNA repair. To tilt the scales in your favor for an HDR edit, you can...

...ll prevent binding to any target protein that is still on the inside of the cell. However, assuming you...hidden epitopes. However, keep in mind that you will still contend with many of the same drawbacks to each ...early steps don’t even involve antibodies, they are still critical to the outcome of your IF experiment and...

...honestly, this still grosses me out. Maybe it’s just me – I didn’t get a smart phone until after I had finished...have used the protocol for this plasmid, and if it still being used. After learning of it's continued popularity...

...receptor (AAVR). Cell lines lacking galactose were still transduced by PHP.B but only if Ly6a was expressed...with PHP.eB, an enhanced version of PHP.B, virus still reached the brain endothelial cells of these mice...capsid variants could unintentionally limit the utility of candidate capsids.  Are you interested in using...

...to recognize my transferable (or “soft”) skills until I was out of the lab and in an office environment...you your next job or promotion. Also, don’t wait until you’re out of the lab to think about these--get ...into science in the first place. Finally, if you still find the idea of not holding a pipette again upsetting...

...nucleic acids. Read on to learn more about this versatile technique.  Immunoprecipitation Overview An immunoprecipitation...typically carried out in one of two ways. Both methods utilize the same basic strategy: immobilize the antibody... One of the most common immobilization methods utilizes bacterial cell wall proteins, Protein A and Protein...

...technique is still used today to test for HeLa contamination. HeLa contamination is still a problem today...You would never thaw any of the Master cell vials until all of the working vials have been used up. This...

...post:  New compact, efficient prime editors A versatile and precise system to improve on-target delivery...10.1016/j.cell.2023.07.039. PMID: 37657419.   A versatile and precise system to improve on-target delivery...lab recently made two deposits that increase the utility of this technique.  By engineering a light-sensitive...

...available to researchers around the world. While we still offer many resources for producing vectors from ...vectors are not as easy to ship as plasmids, they still have a global reach. From 2016–2023, we shared 93,491... Rego, M., Wang, C., Zhang, Y., DeMaio, K. H., Tillgren, M., Fava, A., Clouse, G., Patrick, M., Guerin...

...expressing it from a CAGGs promoter. However FLPe is still less efficient than Cre recombinase. FLPe codon ... Ligand regulated recombinase - keeping Cre out until needed But what can be done when the recombination...Cre/ FLP versions are retained in the cytoplasm until estrogen receptor ligand Tamoxifen is added. Tamoxifen...

...generation of base modifiers available. This system utilizes an RNA template containing the desired edit adjoined...like with base editing, alternative Cas9s can be utilized allowing for PAM flexibility. Fig. 2: Prime...Don’t want to permanently modify the genome but still want to affect gene expression? dCas9 can be used...

...many expression plasmids utilize inducible promoters, which are 'inactive' until an inducer such as IPTG...Arabinose-inducible expression of T7 RNAP; IPTG may still be required for expression F- ompT lon hsdSB (rB...

...the way we apprehend molecular complexes and is still a very useful tool.    Like a guitar hero (that ...SYFP2A, mTurquoise/mVenus and others. CFP-YFP is still one of the best and most used pairs to measure FRET... faster measurement systems, this technique has still many good days and good “gigs” ahead...No offense...

...Escherichia coli One of the most recognizable and utilized model organisms is Escherichia coli. The bacterium... highly characterized genome, and widespread utilization, Arabidopsis has made significant contributions...number of offspring (produces many seeds), self fertilizing, a small yet highly characterized genome, and...

...mutant, this technique uses mutagenic primers that tile across a region of interest, generating multiple...) at a given codon site and multiple primer sets tile across the protein or region of interest. A low ...). If so, the nicking mutagenesis protocol will still work if the orientation of the BbvCI site(s) of ...

...can be achieved through two-protein labeling by utilizing(a) biotin-streptavidin binding and (b) antibody...are protected. Amine-binding viability dyes can still bind free amines on the cell surface, which yields...As a result, the largely negative live cells can still be easily gated on during the analysis. Conclusion...

...track position 5 years after receiving their PhDs, still strive to become academic researchers themselves...universities to hire a glut of trainees at low wages will still be there without restructuring. Positive Points ...highlighting some of the important questions that still need to be answered about the makeup and welfare...

...F Host does (F') or does not (F-) contain the fertility plasmid. A low copy-number plasmid, encodes proteins...hsdR(rk-, mk+) Unmethylated DNA not degraded, cell still can methylate DNA   hsdS(rk-,mk-) Unmethylated...disruption of arabinose metabolism pathway Inability to utilize arabinose as a carbon source leuB β-isopropyl...

...fluorophores, then labeling with the other, you can still record the signal separately for each target.   ...., 2020 multiplexed over 65 targets!) IF is a versatile method with endless possibilities, and there are...Hernandez, J. M., & Germain, R. N. (2020). IBEX: A versatile multiplex optical imaging approach for deep phenotyping...

...normality (or lack thereof). Alternatively, a Quantile vs. Quantile plot can determine if your data deviates...data in ascending numerical value relative to quantiles derived from a theoretical distribution. If your...