We narrowed to 991 results for: igh
-
TypeBlog Post...transposons evolve under contrary selection pressures: high efficiency helps them spread, but also stresses ... evoCAST In the emerging world of gene therapy, highly tailored treatments to specific mutations require...long sequences. In their experiments, PASSIGE had higher efficiency, but evoCAST required less optimization...optimization for new targets and produced a higher-purity product, likely because the integration occurs in a... can’t wait to see where evoCAST takes us on the high seas of gene editing. Bon voyage! References...
-
Plasmids 101: Gene, enhancer, and promoter trapping
TypeBlog Post...proximity to an endogenous gene. Each trap uses a slightly different approach (described in detail below)...for the endogenous gene – its expression levels (high/low) and patterns (tissue/timing) can be monitored...screened. Depending on the marker, traps can be used as high-throughput screens to introduce reporters throughout...the top trap – they don’t require insertion in a highly specific gene region and they don’t disrupt the...Resources References Wood, V., et al., Hidden in plain sight: what remains to be discovered in the eukaryotic... -
Viral Vectors 101: Transductions
TypeBlog Post...are very valuable biological tools, and with the right safety considerations, they aren't dangerous for...transductions can offer benefits such as tissue-specificity, high efficiency, and can be selected to be either integrating...our viral selections to see if we have one that might work for your next experiment. Using transduction...Answering these questions will help guide you to the right method for your experiment. A cool feature of viral...virus can sound daunting, but it is easier than you might think. We have protocols available to get you started... -
What the HEK?
TypeBlog Post...an advantage to use the cell line and we will highlight some of those applications here. Virus production...transfect in bulk and produce recombinant proteins at high levels. In a case where clinical applications of...and if your research doesn’t allow for that, HEK might be the cell line for you. Are HEKs for you? If...you are looking for a human cell line model, HEKs might be a great option! Especially if your research interests... -
Hot Plasmids and Viral Preps - September 2021
TypeBlog Post...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ... TX-TL system is capable of robust expression of high G+C content genes, as validated through monitoring...CRISPR Plasmids and Resources page. Here are some highlights from the past couple months including several... -
Summer SciComm Series: Master of SciComm
TypeBlog Post...yet familiar. The idea that my piecemeal degree might one day foster a meaningful day job only came into...enjoyed a taste of what was to offer, continued straight on to a master's degree in science communication... one day required of me for work, I would have a high level of career satisfaction indeed. For many ...science. Understanding just how the public utilizes insights gleaned from scientific research is, after all...law to poetry. A research discipline in its own right, science communication reflects a necessary confluence... -
FPbase: A new community-editable fluorescent protein database
TypeBlog Post...with a large range of characteristics (color, brightness, photostability, maturation, oligomerization)...choosing a FP, but much of the primary data one might require when making such a decision remains spread...there is a great deal more information that one might like to store about FPs than is currently represented...multi-parametric search queries. Looking for a bright, monomeric FP published in the last few years with... for me? A Practical Approach to Choosing the B(right)est Fluorescent Protein Choosing Your Fluorescent... -
Trading Spaces - Visiting Scientist Edition
TypeBlog Post... 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 ... -
K. phaffii: Rising to the Occasion
TypeBlog Post...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... -
Visualizing Genomic Loci with CRISPR-Sirius
TypeBlog Post...these visualization tools is CRISPR-Sirius, the brightest CRISPR-based tool to date for visualizing genomic...chromosomes in living cells, the system requires high-copy chromosome-specific loci in the genome and ...the gRNA target site. In addition to increased brightness, they also found that the signal to noise ratio...its predecessor CRISPRainbow, CRISPR-Sirius is brighter making it suitable for observing low-copy genomic...copy genes. If you want to visualize genomic loci brightly, give CRISPR-Sirius a try. Find CRISPR-Sirius ... -
Summer SciComm Series: A PhD in Science Communication
TypeBlog Post...grandparents’ medical textbooks by torchlight under the covers. In high school, I went to all the optional...technical jargon that even scientists in neighbouring disciplines might struggle to understand. Why should ...whispered questions from friends and classmates back in high school, and it’s oddly fitting that I’ve ended up... -
New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor
TypeBlog Post...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... -
R Bodies: Membrane-Rupturing Microscopic Tools
TypeBlog Post...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... -
Optimizing Donor DNA for Enhanced CRISPR Genome Editing
TypeBlog Post...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... -
Finding and Joining Your Dream Lab
TypeBlog Post...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... -
How to Deposit Your Plasmids with Addgene
TypeBlog Post.... 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... -
CRISPR 101: Multiplex Expression of gRNAs
TypeBlog Post... 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... -
From Lab Bench to Leadership: Transitioning into Management as a PhD
TypeBlog Post...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 ... -
Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)
TypeBlog Post...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... -
Clear and Accessible Writing
TypeBlog Post... 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...