We narrowed to 842 results for: Eras

...signal you can generate without disturbing the integrity of the membrane, a concern with several of the previously...we will review how these indicators work, their general applications, and why viruses are essential to ...time from signal input to readable output, which generally exceeds that of the VSD GEVIs. However, since ... to the plasma membrane due to the addition of several targeting domains. The quencher and voltage detector...previously described systems. Several adaptations of this method have been to tag, cage, uncage, and trap...your experimental system.   Figure 2: General overview of chemigenetic tools. A genetically encoded...can be used for chemigenetics, biosensors, and generally as biological tools, our blog has you covered....

...have also been using Addgene CRISPR vectors to generate cell lines with knockouts of specific genes. Many...should be able to keep cells moist and hopefully aerated with a thin layer of molecular biology-grade agarose...sterile truncated pipet tips. Indeed, an experienced operator can pick 48 clones from a 10 cm dish in 15- 20...both Hepes and bicarbonate) is brought to room temperature and placed in the hood. The medium from a 10 ...is capped, swirled and tested for the correct temperature by placing on the wrist or cheek. Within less...the agarose cloning method Test your agarose temperature. Cloning cells using an agarose overlay does ...agarose onto the recipient plate at just the right temperature. If the agarose solution is too hot, the cells...

...and use a standard curve and provide some general considerations for scientists planning to use them in ...for the known concentrations (standard curve), generating an equation that allows you to determine the ... triplicate. The closeness of the data points generated by replicate values provides useful information.... With the help of graphing software, you then generate a trendline for the data. The appropriate trendline...users aim for an R2 above 0.95.    Additional considerations When running assays that rely on a standard...To calculate the %CV of the standard curve, one averages the individual %CVs for each point on the curve...assay. Try to minimize these.  Review previous literature or results from labmates to get a sense of the...

...broke down the method into six general steps and outlined the considerations to be made during each step....that occurs depends on the incubation time and temperature, so you may be able to recover good antibody ...fixation*. Organic Solvents Organic solvents, generally acetone, methanol, or ethanol, work by dehydrating...membrane integrity (Fig. 2). For example, saponin interacts with cholesterol in cell membranes and leaves ...Tween-20 are examples of non-ionic detergents that interact with lipids and proteins in membranes and permeabilize...to use your specific antibody for IF, look for general IF protocols for your sample type. Starting with...cross-linking fixation; they require an additional step generally referred to as Antigen Retrieval. I chose not ...

... the Cas9 nuclease in the absence of genomic alterations. These studies utilize a catalytically inactive...dCas9 and targeting gRNA, to identify molecular interactions at this site (Fujita and Fujii, 2013). They ...method can characterize nucleic acid-nucleic acid interactions at a genomic locus (Fujita et al., 2017). CRISPR-mediated...Fujita et al., 2018). Furthermore, the Fujii Lab generated a transgenic mouse line and retroviral expression...when inappropriately expressed can cause facioscapulohumeral muscular dystrophy (FSHD), a disease for ...specific gRNA-dCas9 complex. When combined with next-generation sequencing, this method allows for the recovery...Toshitsugu, et al. "Identification of physical interactions between genomic regions by enCh IP‐Seq." Genes...

...by the Sternberg lab derives from Vibrio cholerae (V. cholerae INTEGRATE - hereafter VchINT) and encodes...and technical considerations Making insertions in bacterial genomes with the V. cholera INTEGRATE system... using Tn5), and as knock-in tools for human therapeutics (e.g. using Sleeping Beauty). Despite their ... CRISPR-Cas, these systems represent the next generation of tools for efficient and programmable insertions...achieved with VchINT, at two different incubation temperatures. Data adapted from Vo et al., 2020. In E...-recombineering, CRISPRa/i, and base editing. Leveraging a unique marriage between distinct biological...

... and several others can be made fluorogenic using environmentally sensitive fluorophores. Overall, self-labeling...student searching couch cushions for spare change, desperate to extract a few more photons from a sample. Chemical...by Roger Tsien, who showed that the selective interaction between a bisarsenical dye ligand (e.g., FlAsH... lipoic acid ligase, biotin ligase, phosphopantetheinyltransferase) – These enzymes catalyze the covalent...corollary to these innovative labeling strategies, several groups—including mine—have been revisiting the ...PMID: 26000849. 2. Grimm, Jonathan B., et al. "A general method to improve fluorophores for live-cell and...Protein Should I Use? Resources on Addgene.org Generate Fluorescent Protein Fusions Request Plasmids from...

...After several decades of trying to address these challenges, researchers have amassed a considerable molecular... overexpress a recombinant protein? It is not generally in a cell’s best interest to overexpress a protein...peptide-based proteosome inhibitors, glutathione S-transferase (GST), which can be fused to recombinant proteins...for affinity and purification An affinity tag, generally a relatively small sequence of amino acids, is...Purification Binding and elution steps use very moderate buffer conditions FLAG DYKDDDD or DYKDDDDK or...interest would eventually get ridiculous (you generally don’t want more tag than protein), 2-3 tags is...IgG-binding domains. TAP has since come to encompass several other tag combinations, though frequently those...

...detected on commercial lateral flow strips, similar to pregnancy tests. With lateral flow strips, the presence... to cleave ssRNA or mRNA. It also exhibits a “collateral effect” of non-specific ssRNA cleavage. How does...sample is first amplified using RPA (recombinase polymerase amplification) or reverse transcriptase (RT)-... for nucleic acid detection almost anywhere as lateral flow strips are easy to transport and work rapidly...diagnostics (e.g. pathogen or virus detection), therapeutics and sensitive genotyping. The beauty of SHERLOCK...Cas12a however recognizes a different PAM site and generates 5’ and 3’ staggered ends after dsDNA breaks. The... DETECTR platform to detect SARS-CoV-2 using a lateral flow strip format. They published this in Nature...

...workflows considerably easier, faster, and more efficient than previous methods. Still, generating reliable...sequence you used. The ICE tool will calculate overall editing efficiency and determine the profiles of...detect or analyze complex edits, such as those generated by multiplexed delivery of multiple sgRNAs. Since...particular parameter was automatically adjusted to generate ICE results. If there are no results or there ... the ICE linear regression is computed during generation of the ICE Score, the Pearson correlation coefficient...around the cut site. It shows, base-by-base, the average amount of signal that disagrees with the reference...sequencing bases downstream of the cut. Summary ICE generates NGS-quality CRISPR editing analysis from Sanger...