We narrowed to 989 results for: IGH@;
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TypeBlog Post...have purified plasmid DNA, this method can be done right in your lab in less than a day. Diagnostic restriction...sequence if necessary. The example plasmid on the right has a total size of 7.3kb, including a 1.2 kb insert...recognition sequence. Most enzyme manufacturers make High Fidelity versions of the endonucleases and/or supply...you to visualize the DNA under ultraviolet (UV) light. Don’t forget to add loading buffer to your digest...
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Sweating the Small Stuff: Details in the Lab
TypeBlog Post...labeling tubes never takes longer than 5 minutes, right?) As a result, I’d always end up stressed and rushing...add in personalized steps like “check drawers for light-sensitive reagents” or “re-sterilize now.” Have ...Moving your reagents from your left side to your right as you use them can help you remember what's been... Fig. 2: Blugene moves reagents from left to right as they use them. Prevent Interruptions If... -
Communicating Your Science With Help From ComSciCon
TypeBlog Post...succeed in communicating. Even the most rigorous, insightful, and novel scientific research will be wasted...your past work has prepared you to contribute at a high level to their team. When you're writing a grant...anyone else in the world to do it. In the lab, it might mean persuasively articulating why the results you've...to be reviewed by interdisciplinary panels. And highly successful scientists need to communicate with ... -
Addgene’s Viral Service - Why Virus? Why Now?
TypeBlog Post...instrumental in helping us establish consistent, high-quality protocols for lentiviral production. Bryan...Janelia Research Campus have provided valuable insight into AAV production. Finally, we are blessed to...surveyed hundreds of scientists about various ways we might expand Addgene’s services. There was an overwhelmingly...helps scientists and allows us to maintain Addgene’s high quality standards?” Under the leadership of Addgene... -
Viral Vectors 101: Inverted Terminal Repeats
TypeBlog Post...sequences are from AAV2 because they demonstrate high promoter activity (Earley et al. 2020), but non-...downstream AAV applications. Tips & FAQs Due to their high GC content and hairpin structures, ITRs are prone...rapidly replicate in a growth culture and result in a high percentage of plasmid DNA with mutated ITR sequences...Next-generation sequencing and Sanger sequencing might fail to correctly read through ITR sequences. GC... -
Viral Vectors 101: Viral Applications
TypeBlog Post...Viral integration of a construct is also reliably higher than traditional transfection and random genomic...transfection method. This is because viral systems have a high infection frequency and the viral titer (the number...responses in addition to antibody response 2) very high immunogenicity 3) and long-lasting immune response...treatment and gene therapy have also been in the limelight. The FDA approved the first oncolytic viral therapy... -
A Deep Dive into BioBuilder
TypeBlog Post...BioBuilder Educational Foundation in partnership with high school teachers to boost interest and understanding...what started in Cambridge with a handful of local high school teachers receiving training modules in synthetic...fall: The BioBuilderClub BioBuilderClub engages high school teams around the world who combine engineering...and STEM-loving students include The BioBuilder High School Apprenticeship, preparing local (Massachusetts... -
REPLACR Mutagenesis: Replacing In Vitro Recombination Methods
TypeBlog Post...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... -
Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers
TypeBlog Post...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 Post...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 Post...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... -
What's New in CRISPR - November 2021
TypeBlog Post... 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... -
Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number
TypeBlog Post...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 Post...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 Post... 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 Post...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... -
Rosella: A Fluorescent pH-Biosensor for Studying Autophagy
TypeBlog Post...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 Post...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... -
Troubleshooting Your Plasmid Cloning Experiment
TypeBlog Post...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... -
All in a Twist: dsRNA
TypeBlog Post...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 ...