We narrowed to 988 results for: IGH@

...mixture of products or from incomplete PCR — although higher DNA concentrations are needed for this method to...retain products with single-stranded overhangs (right branch). The mixed PCR and incomplete PCR methods...with two 5′ overhangs, so these approaches require higher concentrations of insert to succeed. 5′ overhangs... showed that a five-piece assembly reaction was highly efficient (~80%) with 40-bp homology regions. Ten-fragment...method also based on homologous recombination. The higher temperature at which Gibson assembly takes place...

...cognitive biases such as motivated reasoning and hindsight bias, we may fool ourselves and our readers into...replications are unavoidable, but can be minimized with high powered research designs. Unanticipated discoveries...become public eventually). It is typically done right before data collection starts, and should include...example, if you hypothesize that growth rates will be higher in treatment versus control, and you think there...of the study, say a fifth time has unpredictably high rates, should be reported as a new discovery that...

...RNA without altering the genome, but Cas13s have higher on-target specificity. New work from Konermann ...package in low-capacity vectors! These small, but mighty type VI-D enzymes are the latest tools in the transcriptome...including Cas13d-specific regulatory proteins and high targeting efficiency, both of which are described...Cas13d deposit here). Konermann et al.: CasRx is a highly potent, specific editor in mammalian cells Not ...characterized Cas13 enzymes, CasRx also displays very high on-target efficiency; where shRNA treatment produced...

... Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids...here.  Listen to the hot plasmids episode! New, highly efficient RNAi construct for C. elegans gene knockdown...noroviruses can infect millions annually. Because it is highly contagious, it is essential to contain outbreaks...binding experiments. Potential experiments that one might perform using the purified domains include protein...

...is building the software tools for the right people at the right time. To explain a bit further, a software... a strong background of undergraduate research straight to a lab career at a biotech company, I always...operations software for next-gen sequencing (NGS), and high-throughput screening. While some developers had ...about what a software product manager does, you might be wondering what the day to day looks like. Check...

...sensitivity. Wild-type Cas9 exhibits high on-target activity, but also high off-target activity, which may ...sensitivity is the measure of on-target activity (right). Cas9 nucleases that exhibit enhanced site ...et al., 2018) and found that Sniper-Cas9 showed a high degree of both specificity and sensitivity across...Evolved Cas9 variants with broad PAM compatibility and high DNA specificity." Nature 556.7699 (2018): 57. PubMed...

... visitors with information about images that a sighted reader would glean from the image itself.  As we...impaired readers receive the same information as sighted readers.    Figure 1: An image from AAV...image, we thought it would be most helpful to highlight the technical details of collecting the purified... passage of supernatant through each layer. The right panel illustrates a gradient tube before and after...

...surprise then that a lot of our subject matter comes straight from you, whether you realize it or not! Many ...what basic information someone new to antibodies might need to successfully start using them at the bench...bench, along with questions we thought they might have as they go. The guide was then assigned to our science...deeper expertise of our antibody team to rely on. High-impact content like the antibody guide always gets...sketch a western blot out quickly." Final design (right). Immediate and immense regret of the graphic designer...

...does this actually look like? For a selection, you might put cells into a drug-containing media in which ...scientific question can help determine which is right for you. Screen variations Forward and reverse screens...method you choose to assess traits of interest is highly specific to your scientific question. Things to... assay system include: How specific is it? Is it high throughput? Will it be sensitive enough? No one ...screens Selections sound like a great shortcut, right? Who would ever do a screen when a selection can...

...the entire editing system with a simple LED light. Lighting up your gRNA with fluorescent dyes In a scientific... nucleic acids with N-methyl substitutions are slightly more efficient and less toxic to mammalian cells...less than a minute of exposure to the appropriate light. The photoactivatable counterpart to this system...unable to engage a target sequence. Upon a brief light exposure (several seconds) the gRNA is released ...

.... The protein of interest is then eluted using a high imidazole buffer that outcompetes the interaction...protein but not that of the protein of interest. A high imidazole elution buffer disrupts the interaction...concentration of imidazole used in the wash buffer should be high enough to disrupt contaminant-metal ion binding ...therefore more selective. When using systems with higher levels of histidine-containing protein contamination... histidine residues that are present, the more tightly the protein will bind to the metal ion substrate...

...direct approach (left) versus the indirect approach (right). Primary antibodies are represented in blue; secondary... purple; signaling molecules are represented in light green; and proteins are represented in orange. Created...only benefit of secondary antibodies. Finding the right primary antibody for your assay and protein of interest...antibodies are different species and/or isotypes. For highly multiplexed assays, using secondary antibodies ... gel-based method identifies proteins solely by weight and size, relying on the signaling molecule solely...

...research. Congratulations to the winners! Below, we highlight their research and how they will use plasmids ...research into CCDS. The brain and muscles have a high metabolic demand that is powered by ATP. A molecule...bicuspid aortic valve at a rate that is 50-100 times higher than women in the general population,” says Dr.... and can lead to cardiac developmental defects. Eight genes from the X chromosome are expressed at significantly...

...Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids...Chemistry for his contributions to super-resolution light microscopy, comes from his recent Nature Communications...difficult to study in living cells using conventional light microscopy due to their small size (<250 nm), and...probes had to target endosomes, be exceptionally bright (which is necessary for effective STED), and be...

...growth of diverse species. These results further highlight the need for in situ genetic tools for microbiome...Yeast Toolkit by replacing the backbone vector’s high-copy number bacterial ColE1 origin with the RSF1010...microbial population dynamics in the gut can provide insight into how the microbiome responds to infection, ...on the assembly of fluorescent proteins into one bright particle that segregates into one cell during cell...

... and international team assembled the right people at the right time. Be inclusive and seek solutions...Teytelman’s vast network and ability to identify the right opportunities, he was able to bring together talented... email the team. Acknowledgements One of the highlights of 2018 for me was to collaborate with this incredibly...

...conference you’re attending and who approaches you. You might also have cross-team projects within your own lab... errors in our LIMS tracking system. Though you might not have to communicate with a team of software ...document, I update the training frequently. In hindsight, getting a list of frequently used words like ...everyone on the same page. Others in the meeting might also have the same question and asking can help ..., meetings will be shorter and more productive. Higher Productivity. Taking active measures in improving...

...Methods to scale up multiplexed CRISPR systems for high-throughput screening are vastly useful for mapping...CombiGEM uses iterative one-pot reactions with straightforward restriction digestion and ligation steps to... by sequencing its set of barcodes. CombiGEM is highly flexible and can accommodate any genetic elements...Central PMCID: PMC4253859. 4. Hart, Traver, et al. "High-Resolution CRISPR Screens Reveal Fitness Genes and...PMC3972032. 10. Wong, Alan SL, et al. "Massively parallel high-order combinatorial genetics in human cells." Nature...

..., these problems can be overcome by screening a higher number of potential clones. Golden Gate assembly...constructs with an attB recombination scar encoding eight amino acids, but Golden Gate assembly can be designed...that may not be amenable to "domestication", you might want to consider using an alternative method like... for cloning complicated constructs in a single, high-efficiency step. References: Sequential amplification...one pot, one step, precision cloning method with high throughput capability. Engler C, Kandzia R, Marillonnet...

...cell lines to the -omics data. The cell lines with higher expression of the target protein, as determined...expression, are often preferred for this approach. A highly selective antibody should be able to detect even...only be able to detect the target protein at the high levels caused by overexpression. Additionally, if...clones The antibody validation process represents a high level of effort and time, which may not be available... & ABIF Consortium. (2024). A guide to selecting high-performing antibodies for Synaptotagmin-1 (Uniprot....15467 Virk, H. S., Rekas, M. Z., Biddle, M. S., Wright, A. K. A., Sousa, J., Weston, C. A., Chachi, L....