...optimal for experiments requiring high signal to noise ratios, imaging of high frequency glutamate signaling...to create SF-iGluSnFR. This improved variant has higher expression levels in bacteria and produces stronger...properties The improved variants of iGluSnFR3 have higher expression levels and produce stronger fluorescent...Biosensors A Practical Approach to Choosing the B(right)est Fluorescent Protein Visit the Fluorescent Protein...

... be ideal for you. Here, we will chat about who might want to explore this role, how to seek out such ...commitment, planning, and funding – they may not be right or necessary for many scientists! Unless some of...from third parties or your home institute. Discuss rights Do you expect to generate data that will contribute...It is never too early to discuss authorship and rights for anything that could be generated during the...opportunity, but also a really good time! If it sounds right for you, start thinking outside of your current ...

...buck - it grows at higher cell densities than most of its counterparts, yielding higher production power... phaffii in the 1970s with the goal of producing high protein animal feed. Due to increasing prices of...flask volume. What’s even better? K. phaffii is highly efficient at protein secretion, making harvesting...knock-in or knock-out) is essential to your work, you might want to pick a yeast strain best suited to it. Putting...

...figure to the right), we identified R-body mutants that extend at both lower and higher pH. We initially...putting the operon under a strong promoter results in high R-body expression  (Schrallhammer et al 2012). This...environmental conditions. More speculatively, R bodies might endow divergent organisms with new functions. For...understanding of how R bodies assemble and function, we might be able to endow cells with new ways to interact...

...sides of the break, we determined that Cas9 held tightly to three of the four strands of the cleaved duplex... the PAM-proximal side, consistently supported higher frequencies (up to 60%!) of gene editing than single...have shown that this technique supports extremely high frequencies of genome cutting and, when donor DNA...included in the nucleofection reaction, extremely high frequencies of HDR. The detailed protocol for nucleofection...Central PMCID: PMC4383097. 3. Kim, Sojung, et al. "Highly efficient RNA-guided genome editing in human cells...

...In a big lab, the advisor might not be around much and your real boss might be a postdoc – this isn't ...competitive style to spur you to succeed or you might enjoy working closely with others to accomplish ...before you take the position. Am I choosing the right mentor? Make sure your advisors will be good role...candidate to ask lab heads, lab members and lab neighbors during the interview process. What if I want...

... you to select cells with high GFP expression. These cells have the highest levels of Cas9 and gRNA expression... which method you use, the procedure will vary slightly.   Two Golden Gate options that are available...expression, and thus the highest frequency of genome editing events. Yamamoto Lab Multiplex CRISPR/Cas9.... “Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing.”  Nature Biotechnology...al. “A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and...

.... If submitting plasmid(s) using a spreadsheet (right), click the “Download Deposit Spreadsheet” button..., and plasmid description/experimental purpose (right). On the article page, enter the name, type, and...The more sequence data available, the better. We highly encourage you to upload full sequence data whenever...cannot be propagated in DH5α. For plasmids with highly repetitive sequences (which can be prone to recombination...Figure 12: The "Copy" button is located near the top right of each data entry page. Once you have completed...necessary by my institution or other third party rights holders. Once this process is complete, you will...

...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...

...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...

...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ...close proximity to the GFP-tagged protein (albeit slightly offset due to the nanobody-GFP interaction). The...the AAV capsids (left) and transduction patterns (right).  Both AAV-PHP.eB and the AAV.CAP-B capsids are...mammalian cells. Long sequences traditionally have higher risks of off-target mutations. These plasmids contain...

...to induce high levels of gene activation for both reporter and endogenous genes. It is highly specific ...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ...target only requires a small epitope tag (FLAG is eight amino acids, HA is nine), multiple repeats of the...

...hybridoma, you can choose one that produces high-affinity/high-avidity antibodies. The same hybridoma is...drive can be identified by sequencing the heavy and light chains. To circumvent any possible genetic drift...  Figure 2: Changes to the heavy chain or light chain genes within a hybridoma impacts the specificity...validating monoclonal antibodies, their cost is higher than that of polyclonal antibodies. For example...

...realized that working in the wet lab wasn’t the right path for me. I began exploring alternative career...track and feel supported. Another team member was highly self-motivated and excelled when given the freedom... made similar transitions can provide you with insights, lessons from their experiences, and practical...perspectives and facilitated a team meeting to highlight their complementary strengths. By encouraging ...

...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...

...Since its early years, the light microscope has been a powerful tool in almost all disciplines. In biology... cells, has become an established alternative to high-priced commercial devices.     Fig. 2: Workflow...Workflow for open software (left) vs. open hardware (right).    Taking this all together, the essential...open hardware. Upper left panel shows the path of light through a microscope. Lower left panel shows an ...differentiating monocyte with planes and hardware to it. Right panel shows workflow of open hardware.    Conclusion...

... 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...