...the pandemic; for more discussion on that, see Pyhtila et al 2023.)  One of our favorite observations ...CRISPR Plasmids by Addgene,” in The CRISPR Journal (Pyhtila et al., 2023). Our thanks to Rodolphe Barrangou...publications.  References and resources References Pyhtila, B., Kasowitz, S., Leeson, R., & Barrangou, R. ...

...Chemigenetic sensors take a hybrid approach and utilize both a genetic component as well as a small molecule...fluorescence all have room for improvement. These sensors still have many useful applications, though, and if you...this two-part fluorescent system, and rhodopsin still functioning as the voltage sensor. FP-rhodopsin ...

...across multiple model organisms (3) lies in the versatility of its delivery methods. Researchers can conveniently...unhappy with the new method and 65% wanted to wait until someone else optimizes an NgAgo protocol for their...complex with a 5’-P-ssDNA guide at room temperature. Until that time, those who have been lucky with NgAgo ...

...done the technical design of your blot, but are still finding yourself having issues, you’re in the right...blocking conditions on the spec sheet. If you’re still seeing high background after following manufacturer...yourself. But even with a validated antibody, it can still take time for you to get it to work well. Dilutions...antibody dilution. Adjust the antibody dilution until your protein dilutions are in the dynamic linear...

... the base of the stem. This buys you a few days until new inflorescences begin to grow. This will also...too old to transform. This transformation could still work, but the efficiency will be low. Tips: Use...plants: If you transform early, your plants will still have plenty of time to produce seeds after you expose...

...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...

...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...

...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...

...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...

...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...

...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...

...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 ...

...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...