We narrowed to 989 results for: IGH@;
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TypeBlog Post...ancestry groups rather than self-reported race, it highlights the myriad issues researchers need to reflect...associated dependencies across ancestry groups (right). Seeking an explanation First, they looked...guides from being designed that target regions with high genomic variability. Moving forward It took a ...Stephanie M. Fullerton et al. Getting genetic ancestry right for science and society." Science 376, (2022): 250...
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CRISPR Antimicrobials
TypeBlog Post... may be just the method needed to jumpstart the fight against antibiotic resistance. Advantages of CRISPR... despite this resistance gene being encoded by a high-copy plasmid. In the case of the genomically encoded...distinct beta-lactam resistance genes, again with very high specificity. This double phage system both favors...bacteria. In mixed populations, their phages were highly specific for certain bacteria, just like the CRISPR...new strategy can help us win back ground in the fight against bacterial superbugs. References 1. Citorik... -
Cpf1: A New Tool for CRISPR Genome Editing
TypeBlog Post... species, Zetsche et al. hypothesized that Cpf1 might represent a functional CRISPR nuclease that could... cleaves relatively far away from the PAM, NHEJ might retain the PAM site. Therefore, if HDR does not ...possibly mediate HDR. This “second chance” mechanism might improve the frequency of desired HDR edits, but ...editing efficiency for the Cpf1 orthologs was only slightly lower than that of the widely used SpCas9 and ...that AsCpf1’s off-target rate is similar to that of high fidelity Cas9s eSpCas9 and SpCas9-HF1. Both AsCpf1... -
The Importance of a Fun Workplace: Company Culture at Addgene
TypeBlog Post...has drastically increased, it would have been a tight squeeze for all of us to fit in Joanne’s back yard...discovered my hidden talent in Ladder Ball... there might be a possibility it was just beginner’s luck, but... (only if it’s cool with your legal department) right in your breakroom. 5. Throw some physical activity...you have a very intricate itinerary, there is a higher likelihood that something will go awry. Don’t plan... My grandma used the old adage “many hands make light work” all the time. She was a wise lady for many... -
Fluorescent Proteins 101: Green Fluorescent Protein (GFP)
TypeBlog Post...shielded from quenching by aqueous solvent. This tightly-packed structure explains the importance of the...permutations within each color also exist with only slight variations separating them. Please note that many... Improves folding at 37C, reduces aggregation at high concentrations T203Y Red-shifts the wavelength...biosensors here. Optogenetics: Scientists can use light to detect, measure, and control molecular signals... blog, you should! He's got great articles and insight on the newest tools and techniques in fluorescent... -
CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More
TypeBlog Post... This powerful negative selection system ensures high editing efficiency; the only non-edited cells to...The simpler system of pCRISPomyces-2 displays a higher editing efficiency, perhaps due to its condensed...Streptococcus thermophilus, Cas9sth1 as a robust and highly active enzyme for transcriptional silencing Mycobacterium...efficiency of knockdown and developed new, more tightly controlled, Tet responsive promoters to avoid leaky...dCas9 variant as well. Furthermore, they provide a high-fidelity (HF) variant of the BE3 that further improves.../10.1038/nbt.3011 Cobb RE, Wang Y, Zhao H (2014) High-Efficiency Multiplex Genome Editing of Streptomyces... -
Analyzing CRISPR Editing Results with ICE from Synthego
TypeBlog Post...found that the accuracy of ICE analysis results was highly comparable (within r2=0.96) to that of NGS data...coefficient (r) is also computed and reported. The higher the R2 value, the more confident you can be in ...the Download Analysis Data button on the bottom right of the analysis screen. ICE analysis details Contributions...genomes that contain the indel. The discordance plot (right panel) shows the level of disagreement between the...that continues after the cut site, representing a high level of sequence difference. Traces The Traces... -
AAV Titers: Where Do They Come From And What Do They Mean?
TypeBlog Post...SDS-PAGE...Electron microscopy… Any guesses? That's right... ways to titer AAV! The point is there are many...linear region of the standard curve. In addition, high and low-concentration samples can also behave differently... reference sample of known titer, which is at a higher titer than the AAV RSM. By using these samples ...) AAV titers are approximations and they depend highly on the experimental conditions used, titer values...PMID: 10455443. 3. Hindson, Benjamin J., et al. "High-throughput droplet digital PCR system for absolute... -
Save Time with Transient Plant Leaf Transformations
TypeBlog Post... procedure (Mortensen et al., 2019). Our method might provide some useful guidelines for the development...transformation was successful at all. Once we were on the right track, we switched to luciferases to more easily...simple and easy technique to implement. Success might vary between experimenters. Vacuum infiltration ...especially problematic with reporter genes as you might observe expression from Agrobacterium, but not the...leaf transformations, but we hope that these tips might help to get you started. Once you have established... -
Plasmids 101: Gateway Cloning
TypeBlog Post...technology has become very popular as a rapid and highly efficient way to move DNA sequences into multiple...two new recombination sites (attL-left- and attR-right-, Figure 1). Under certain conditions, the attL ...’ll walk through an example experiment where we might use Gateway cloning to generate our desired constructs...another, the inserted DNA fragment stays in frame. High efficiency The positive (antibiotic) and negative...M. (1992). A Genetic Switch: Phage (Lambda) and Higher Organisms (Cambridge, MA: Cell Press). Additional... -
Antibiotic Resistance: An Old Solution but a New Problem
TypeBlog Post...three scenarios, but a short treatment regiment might wipe out bacteria that are completely susceptible...concentrations of a drug, but will be killed by a higher concentration, or sometimes more exposure time ... corresponds to the level of susceptibility you might expect if a certain antibiotic were chosen for treatment...survive for millennia. What we can do, however, is fight smarter, not harder. We can utilize tactics like...antimicrobials to learn how scientists are using CRISPR to fight bacteria. From a societal standpoint, we can all... -
Changing Labor Laws Bring Increased Postdoc Wages
TypeBlog Post...Department of Labor’s general guide to the effect on higher education, you can see that those in teaching roles...lot of us, frankly quite surprising. Consortia of higher education institutions submitted letters to the...exemption. Discussions about the drastic effects on higher ed had been seized upon by opponents of the ruling...convincing as the recommendations for postdocs to be on higher salaries (which have been made by multiple organizations...recommendations that postdoc salaries be raised to $50,000 - higher than the level they will be raised to on Dec 1st... -
5 Reasons to Use Reddit for Science Communication
TypeBlog Post...you can see some of the stats from the AMA to the right - the Reddit community is active, engaged, and has...resources at her disposal, she was writing all day and straight on through her designated two hours for answering... to other scientists who will find it useful, I highly recommend doing an AMA! Before I turn it over to... Kudos to the moderators for keeping the quality high and thus enabling a productive discourse. They even...to resources. I’ve said this before, but I also highly recommend taking every opportunity you get to speak... -
Viral Vectors 101: Pseudotyping
TypeBlog Post...scientists achieve a high titer (reviewed in Joglekar and Sandoval, 2017). This high titer is one of the...your virus, there are many other reasons when you might pseudotype your virus: Lowering cytotoxicity. ...reaching the target cells. VSV-G is also toxic at high concentrations as described above and is inactivated...vector to be transported retrograde and also has higher neuronal transduction efficiency and a better safety... -
Finding nucleic acids with SHERLOCK and DETECTR
TypeBlog Post... develop and optimize the method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK... the promiscuous cleavage and degradation of neighboring ssRNA and ssDNA by Cas13 and Cas12a, respectively... of CRISPR-based diagnostics. SHERLOCK- Specific High Sensitivity Enzymatic Reporter UnLOCKING Cas13 (...SHERLOCKv2 provides a method to detect nucleic acids with high sensitivity and specificity without compromising...the ability to rapidly detect nucleic acids with high selectivity and sensitivity from patient samples... -
Harnessing Bacterial Toxins for Allelic Exchange
TypeBlog Post...created vectors for Lambda Red that contained one of eight bacterial toxin genes isolated from E. coli or Psuedomonas...both rhamnose- and arabinose-inducible promoters. Tight control of toxin expression is absolutely necessary...induction system is well-characterized and quite tight (Giacalone et al., 2006). The Chen lab demonstrated... selected the three toxins so that at least one might be useful in a broad array of pathogenic Proteobacteria...expression in Escherichia coli using a rhamnose-based tightly regulated and tunable promoter system." Biotechniques... -
Viral Vectors 101: Integrase-Deficient Lentiviral Vectors
TypeBlog Post...cells. However, for some applications, integration might be considered a double-edged sword with unintended... lentiviral vectors, how they work, and why you might choose them over integrating lentiviral vectors....lentiviral vectors? There are several reasons why you might choose an IDLV over a lentiviral vector: 1. No risk...mind is that titering these IDLVs may require a slightly different protocol than regular lentiviral vectors...Reyon, D., Joung, J. K., & Sander, J. D. (2013). High-frequency off-target mutagenesis induced by CRISPR-Cas... -
Multiple Plasmids at a Low Price: Kits from Addgene
TypeBlog Post... together on a tight budget. To help popularize our kit resources, we’re highlighting just a few of ...covering a wide range of fields from Addgene. What you might not know is that Addgene distributes curated collections...Morgan’s blog post on the PUF Assembly Kit for some insights on its potential from its creators. We hope you... -
3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells
TypeBlog Post...change and screen a few hundred clones to find the right one Step 2 – Edit the cells a second time using ...corrects the blocking mutation and again identify the right clone from a few hundred. From these two steps, ... few hundred clones - Still better than 10.000, right? (Kwart, Paquet, et al 2017). Find Additional CRISPR...increasing mutation incorporation efficiency, we might exploit this relationship to direct the zygosity...Additional Resources on the Addgene Blog Learn about High Fidelity Cas9 Variants How to Design Your gRNA Delivering... -
Antibodies 101: Beyond Surface Labeling
TypeBlog Post... cell mixes, like bulk lympho-/splenocytes, you might only be interested in one of the various cell populations...markers of all cell types except CD8+ T cells. We might choose the markers CD4, CD19, and CD11c, which would...rest can be excluded, or “dumped”. From left to right: B cell, CD4+ T cell, dendritic cell, CD8+ T cell...host species reactivity. When sorting cells with a high resolution for specific downstream applications,...immunity. When he's not in the lab, Paul enjoys weight lifting, sailing, and reading. More resources...