We narrowed to 839 results for: IGH@;
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TypeBlog PostPublishedFeb. 10, 2016, 3:30 p.m....disease. CRISPR has made genomic SDM relatively straightforward, but plasmid-based SDM has lagged behind. While...well, but poor template digestion can result in a high number of background colonies carrying the unmodified...plasmid is not maintained. Recombination efficiency is high enough to negate the need for selection usually ...introduce mutations. Using commercially available high-fidelity polymerases such as KOD and Phusion, which...
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Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers
TypeBlog PostPublishedJuly 28, 2015, 2:30 p.m....Mendenhall: The benefits are two-fold: CRISPR/Cas9 offers high efficiency of inducing double strand breaks and ...weeks. CRISPR saves you a couple of months. It’s higher throughput and faster. It’s also easy to run things...transcription factors, but there are many reasons you might want a system like this. It’s actually surprising...with this antibody problem. We thought it was the right thing to do to put these out there. It’s a simple... -
Will You Be My Mentor? Finding and Asking for Mentoring Support
TypeBlog PostPublishedFeb. 4, 2014, 3:50 p.m....interested in following? Someone you admire for their insight and way of expressing it in meetings? The more ...advice. Your family and friends know you well and might have good ideas for who you should connect with....cautious and make sure to choose one that comes highly recommended from someone you trust. A good coach...compliment to ask someone to share their wisdom and they might just not have the time. Thank them anyway and move... -
CRISPR/Cas9 FAQs Answered!
TypeBlog PostPublishedMarch 13, 2014, 4:08 p.m....that wildtype Cas9 with optimized chimeric gRNA has high efficiency but has been shown to have off-target...prefers a 'G' at the transcription start site to have high expression, so adding this G could help with expression... culture unless the targeting efficiency is very high. Q6: How should I design experiments for multiplexed...but the efficiency will be lower, although this might still be okay if you are introducing a selection...10.1016/j.cell.2013.08.021 Ran, F. A., Hsu, P. D., Wright, J., Agarwala, V., Scott, D. A., & Zhang, F. (2013... -
Rosella: A Fluorescent pH-Biosensor for Studying Autophagy
TypeBlog PostPublishedApril 13, 2017, 2:30 p.m....neutral pH compartment, like the cytosol, to the higher pH of the vacuole. Read on to learn more about ...enzyme. Pho8 phosphatase activity or the molecular weight shift of Pho8 from its uncleaved to cleaved form... dual color-emission biosensor named after the brightly-coloured Australian parrot. It’s comprised of ...Oligomerization Seeing Red: Simple GFP Photoconversion Light Sheet Fluorescence Microscopy Additional Resources... -
CRISPRainbow and Genome Visualization
TypeBlog PostPublishedFeb. 28, 2017, 3:30 p.m....cyan, or magenta light. A gRNA with all 3 distinct hairpin sequences produces white light. Created with ...from red, blue, and green channels overlaid on a bright-field image. Image from Ma et al, 2016. Using... A gRNA with all three hairpins generates white light. All 6 gRNAs can be expressed in a single vector... -
Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number
TypeBlog PostPublishedSept. 4, 2018, 12:53 p.m....concentrations. Higher IPTG concentration leads to higher RepA expression which results in high plasmid copy...repressor because TALEs can be programmed to bind tightly to arbitrary DNA sequences and have been shown ... -
RaPID Detection of RNA-protein Interactions
TypeBlog PostPublishedAug. 16, 2018, 12:40 p.m....interactions by cross-linking RNAs and proteins with UV light, followed by an antibody pulldown of a protein of...for Affinity Purification (CRAPome), to identify highly biotinylated background proteins reported by other...and identified QKI, an RNA-binding protein that’s highly expressed in neural progenitor cells. Knock-down...Response Element (IRE) RNA motif of the ferritin light chain RNA transcript. IRE motifs are bound by IRE... -
RANbodies: Reporter Nanobody Fusions
TypeBlog PostPublishedApril 10, 2018, 12:56 p.m.... is weaker than reporter staining. Picking the right reporter for your RANBody While all three reporters... fluorescently labeled tyramide molecule into a highly reactive radical, which is then covalently linked...antigens. Spaghetti Monster Spaghetti Monsters are highly antigenic tags that have 10 HA or 10 MYC tags built... plasmids from Penn State Need help picking the right fluorescence microscopy technique to use? Check ... -
Oh, The Places You Can Go: Careers in Science Communication - Writing for a Research Institute
TypeBlog PostPublishedFeb. 2, 2018, 3:20 p.m....feels she was lucky to have been in the right place at the right time when she started her scicomm career...“There were jobs that I didn’t seem to have the right skills for in terms of PR types of things or communications...leverage unpaid work for paid work: “If you use it right, it could help you get your name out for stuff that... -
Troubleshooting Your Plasmid Cloning Experiment
TypeBlog PostPublishedSept. 24, 2019, 12:55 p.m....result in any cells, or all the colonies screened might not contain the correct plasmid. There’s a lot that... 24 bp with the template and if the sequence has high GC or AT content, increase the overlapping region...Full lawn of cells. The antibiotic in your plate might not work. Check the plates! Streak on them an E....t get any positive clone after these steps, you might want to reconsider your approach, and try a different... -
What's New in CRISPR - November 2021
TypeBlog PostPublishedNov. 18, 2021, 2:15 p.m.... edition of “What’s New in CRISPR” - where we highlight a few of the newest CRISPR plasmids available ...cells. The “V4” version (named CasMINI) had the highest activity for gene activation and base editing,... not modify the Cas12f1 enzyme. The result is a highly efficient Cas system with very compact genome editors... -
All in a Twist: dsRNA
TypeBlog PostPublishedFeb. 2, 2023, 2:15 p.m....RNA! Double stranded? Well, that’s just for DNA – right? RNA performs almost all of its biological functions...replication process. dsDNA viruses yield dsRNA through a slightly different mechanism thought to be associated with...always around. R-loops are particularly enriched in highly transcribed regions and repetitive regions, such...RNA:DNA hybrids levels are allowed to reach very high or low levels, possibly due to dysregulation of ... -
Addgene's Tips for Plasmid Quality Control
TypeBlog PostPublishedJan. 14, 2016, 3:30 p.m....plasmid verification, and using NCBI BLAST. Tips for high-throughput plasmid sequencing If you’re like us,...Sanger sequencing is done from glycerol stocks (overnight cultures with 15% glycerol), as opposed to purified...handling of large numbers of samples. When we grow overnight cultures for glycerol stock production and storage...can make your life much easier by selecting the right primer. This often starts by determining what features... -
Popular Retroviral Vectors and Their Uses in Scientific Research
TypeBlog PostPublishedJuly 24, 2018, 12:51 p.m....systems Gamma-retroviral vectors have a relatively high transduction efficiency, but in contrast to lentiviral...small-molecule libraries and can be combined with high-throughput sequencing to perform multiplexed analysis... cassette. It can be used to produce a ~10-fold higher titer of lentiviral particles than the original...lentivirus provides significant genetic power and allows high throughput genetic modulation studies in mammalian... -
A Guide to Designing a Scientific Poster: Content, Layout, and More
TypeBlog PostPublishedOct. 22, 2019, 12:20 p.m....acknowledgements sections. Some other additions you might wish to include are a references section, contact...navigate the poster from column to column left-to-right, which is the natural way that people expect to ...illustrations above) so that the color fades to a lighter or darker version of itself from the top of the...sure that the color of your font is compatible. Use high-contrast between these colors so that your text ... -
Plasmids 101: The Wide World of Natural Plasmids
TypeBlog PostPublishedOct. 8, 2024, 1:15 p.m....just for lab specimens — there is evidence that lightning can also promote gene transfer in soil bacteria...microbial offense One way to triumph over your neighbors is to eliminate the competition, and Penicillium...specialized degradative capabilities are often tightly regulated to be active only in the presence of ...Simonet, P. (2001). Laboratory-scale evidence for lightning-mediated gene transfer in soil. Applied and Environmental... -
Making CRISPR Plasmids Using Fragmid
TypeBlog PostPublishedAug. 29, 2024, 1:15 p.m....If you’ve ever been looking for just the right CRISPR vectors on Addgene and found instead ones that ...Doench reports even the greenest members of his lab, high school interns, were able to use Fragmid to design...making it easier to get the vectors that are just right for your experiment. As always, we recommend checking...components by clicking the blue text in the upper right-hand corner. Step 3: Reviewing the ... -
Design Tips for Prime Editing
TypeBlog PostPublishedJan. 23, 2025, 2:15 p.m....location bind to Cas9 with canonical sgRNA structure (right). Created with BioRender.com. What to edit The...3 or more mismatched bases can often evade MMR (right). Created with BioRender.com. For specific prime...prime editors bound to sgRNAs only drive nicking (right). Start with nicking sgRNAs ~50 bp from the prime...2024). Figure 9: epegRNAs (left) and PE7 (right) offer two different strategies for protecting pegRNAs... -
CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9
TypeBlog PostPublishedNov. 14, 2024, 2:15 p.m....genome is made of protein-coding genes, and you might be wondering what the rest of our genome could possibly...non-coding RNAs! You probably didn’t see that coming, right? Non-coding RNAs (ncRNAs) are transcribed but do...versatile option. It can be used to target lncRNAs with high specificity, particularly in applications requiring...Hazan J, Bester AC. CRISPR-Based Approaches for the High-Throughput Characterization of Long Non-Coding RNAs...