...reproducibility. Cause 1: Inability to repeat previously performed experiments Researchers should be able to follow...thorough notes on how lab procedures were actually performed, potentially curtailing future confusion (and ...previously published results. This over-reliance on the perfect story for a “high impact” publication can lead ...situation in Figure 1 where many scientists are performing similar experiments. Biology is messy enough ...practices that fall into this category include performing experiments with too few replicates leading to... an experimental design to a journal prior to performing the experiment (or set of experiments), the design...

...Fluorescent proteins (FPs) offer scientists a simple yet powerful way to tag cellular proteins and investigate protein...scientists later developed red fluorescent PA-FPs by performing several rounds of random mutagenesis screening...most scientists working on in vivo imaging models perfer the PA-RFPs over PA-GFP. Another class of irreversible...fluorescent protein for your experiment Similar to performing experiments with standard FPs, before choosing...characteristics, as the monomeric state is less likely to interfere with endogenous protein structure and function...before your experiment even begins! If you are performing multicolor experiments, you will want to be sure...

...early applications of CRISPR/Cas9 Nonetheless, as powerful as CRISPR-Cas9 has been in the lab and as promising... more. The downside of CRISPR 2.0? It can only perform one function at a time because, in most cases, ... we consider CRISPR 3.0: Casilio. Casilio is a powerful platform that allows CRISPR/Cas9 to multitask ...labeling of telomeres and centromeres. Another powerful potential application is the assembly of stoichiometrically...with a research focus on engineeringbetter, more powerful ways to investigate the genome, epigenome, andtranscriptome...

...convey your ideas clearly and strategically is powerful in almost any career. It allows you to share and...goal.   As a scientist, public speaking isn't a performance art. You’re (usually) presenting either to share...if I need to,” or “It is okay if it doesn’t go perfectly,” can help. (And do remember to breathe!)   Public... exceptionally difficult for you or not going perfectly, you truly are a rock star.  The best thing about...in its most difficult form, which makes it the perfect transferable skill. Almost every other form of ...

... to be recruited per targeted gene. Performance SunTag performs better than first generation activators...elongation, depending on which sequence is targeted. Performance dCas9-VP64 activation is generally thought of...additional activation domains (HS1 and p65). Performance When targeting single genes, SAM consistently...in tandem to recruit transcription factors. Performance Generally, VPR is found to have significantly...

...do anything. Viruses are complicated and they perform complex biological tasks, both of which are factors...you’re unsure about the integrity of your virus, perform a diagnostic restriction enzyme digest and compare...transfected packaging cells. This check can be performed routinely and without disrupting the flow of your...viral vector. This antibiotic selection can be performed approximately 72 hours after transfection and ...Another method of ensuring viral production is to perform PCR against components of the viral vector using...

...even more.) However, none of the tools we found performed batch guide design, batch primer design or batch...transformation (illustrated in the graph below) is performed because weak gRNAs (CFD < 20) bind almost not ...strong gRNAs (CFD > 80) bind almost as much as perfect gRNAs (CFD = 100). CFD scores (which range between... cut and the insert, the more poorly the guide performs HDR. Optimal mutation in-depth One the most sophisticated...donor RNA Design primers Review and order reagents Perform your wet lab experiment Analyze the data For more...

...leaves a “scar” in the chromosome, sometimes interfering with expression of surrounding genes. However...and Prather, 2015). Both of these systems are powerful, but they rely on exogenous enzymes, require multiple... markers for allelic exchange. To do this, we performed the following modifications to pDS132 (Philippe...Hospital where he is now an attending physician. He performs post-doctoral research in the lab of Matthew Waldor...PMC3748948.  Khetrapal, Varnica, et al. "A set of powerful negative selection systems for unmodified Enterobacteriaceae...

...status as a Biosafety Level 1 microbe makes it a perfect candidate for introductory teaching and research...generation time of around 20 minutes, they are perfect for molecular cloning and metabolic engineering...a model organism has its perks, it is far from perfect. One of the most limiting attributes of rats is...Zhang, C., & Xing, X.-H. (2018). Pooled CRISPR interference screening enables genome-scale functional genomics...genomics study in bacteria with superior performance. Nature Communications, 9(1). https://doi.org/10.1038...

...Camera 4K. It’s wonderful and simple to use, but when you need to juggle filming and performing on camera, ...laptop raised on an ironing board, giving it the perfect height (Fig. 2). We recorded video using a simple...celebrating Massachusetts Nonprofit Day, which was the perfect opportunity to test shooting over Zoom, everyone...talent could just focus on their interview or performance. By this point, towards the end of the summer...

...use a unique and powerful high-throughput screen to select variants based on performance metrics specifically...Near-infrared (NIR) fluorescent proteins promise better performance for in vivo use and deep-tissue imaging. Vladislav...for screening candidate biosensor variants for performance under two-photon fluorescence lifetime imaging...produce a temperature sensor that significantly outperforms previous designs. B-gTEMP is excitable under ...

...use. Or you can really capitalize on the dye’s performance and use super-resolution or single-molecule techniques...fluorescent ligand and see the results right away. Or perform pulse-chase experiments by sequentially adding ...priority, SNAP-Tag or HaloTag have been shown to perform differently for different ligands (Wilhelm et al...super-resolution imaging, HaloTag gives better performance than SNAP-tag (Erdmann et al., 2019). Check out...new ligands for these systems, enhancing their performance and ease of use. With so many possibilities from...

...phenotypic readout, similar mapping could be rapidly performed by targeting all potential components of a signaling...antibodies. Antibodies could further be used to interfere with the identified surface proteins, and such...available through Addgene. This means you could perform a screen with one library from Addgene, and then... in vitro, but there is enormous potential in performing in vivo screens, studying genes affecting cellular...the information that you have input in the web interface, making it convenient for you to later identify...

...fluorescent biosensor. Image from Chesterfield et al., 2020     Chesterfield et al., ACS Synthetic Biology... and physiology in vitro and have emerged as a powerful model for understanding human physiology, development...the Rawlins Lab have compiled a new toolkit for performing genetics studies in organoids. The Organoid EasyTag...

...format not only enables you to discuss a person's performance or what has been learned, it helps to plan effective...of indirect language You must be honest about performance problems. While giving corrective feedback isn... or use indirect language to talk about real performance issues. Watch this little video to learn more...don’t know how you actually feel about their performance. Giving honest and helpful feedback gets easier...

...unwind the DNA and check for sequence matches; a perfect guide RNA will do you no good if there is no adjacent...tolerates mismatches in the next 8-10 bases after perfect pairing in the seed sequence.   Figure ...lab mutated the residues in this loop to produce SuperFi-Cas9, a high-fidelity variant that still retains... CRISPRing!   Molecular graphics and analyses performed with UCSF ChimeraX, developed by the Resource ...Bravo, J. P. K., Liu, M.-S., Hibshman, G. N., Dangerfield, T. L., Jung, K., McCool, R. S., Johnson, K. ...

...primers that contain the regions of homology. A powerful resource for planning an NEBuilder HiFi DNA Assembly... which is freely available online. Its simple interface can guide you through both experimental and primer...design results in covalently joined, scarless DNA – perfect for transforma­­­tion or as a PCR template. Some...designing primers with your desired mutation, performing PCR to generate the mutation and linearizing ...

...We all know that yeast is a powerful eukaryotic model organism – its genome is easy to manipulate, it...expression   K. phaffii first gained attention for its powerful promoter of a gene important for methanol usage...essential for sequestering certain enzymes and performing specific oxidative reactions. They are found ...predominantly uses non-homologous end joining (NHEJ) to perform DNA double strand break repair instead of homologous...

...constructs Anti-GFP [N86/8R] ready-to-use antibody Hyperfolder YFP CRISPR-Csm: A novel RNA targeting system...Another reason is that some antibodies may just not perform as well in one application as another, so it is...eLife.43322. PMID: 30667360; PMCID: PMC6377228. Hyperfolder YFP- a chemically stable tool for microscopy ... The Liu and Petsko labs recently engineered hyperfolder YFP (hfYFP), a FP well-suited for the demands...

...of that nucleotide. At that position, T4 would perform the favored polymerase reaction and subsequently...annealing. Elledge realized that he could generate imperfect “recombination intermediates” through PCR and ... To overcome this limitation, one option is to perform a hierarchical assembly, assembling fragments in...Choosing a Molecular Cloning Technique Choosing Your Perfect Plasmid Backbone   ...