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Showing: 541 - 560 of 987 results
  1. A Deep Dive into BioBuilder

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    Blog 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...
  2. Addgene’s Viral Service - Why Virus? Why Now?

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    Blog 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...
  3. Communicating Your Science With Help From ComSciCon

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    Blog 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 ...
  4. Sweating the Small Stuff: Details in the Lab

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    Blog 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...
  5. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

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    Blog 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...
  6. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

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    Blog 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...
  7. Will You Be My Mentor? Finding and Asking for Mentoring Support

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    Blog 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...
  8. CRISPR/Cas9 FAQs Answered!

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    Blog 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...
  9. All in a Twist: dsRNA

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    Blog 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 ...
  10. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

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    Blog 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...
  11. CRISPRainbow and Genome Visualization

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    Blog 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...
  12. RaPID Detection of RNA-protein Interactions

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    Blog 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...
  13. RANbodies: Reporter Nanobody Fusions

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    Blog 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 ...
  14. Troubleshooting Your Plasmid Cloning Experiment

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    Blog 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...
  15. What's New in CRISPR - November 2021

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    Blog 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...
  16. Addgene's Tips for Plasmid Quality Control

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    Blog Post
    ...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...
  17. Design Tips for Prime Editing

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    Blog Post
    ...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...
  18. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

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    Blog Post
    ...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...
Showing: 541 - 560 of 987 results