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We narrowed to 741 results for: REV

Showing: 181 - 200 of 741 results

  1. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...In this study, the labs found that anti-CRISPR prevents Cas9 activity in HEK293 by reducing editing from... Acr samples. Acr proteins can also be used to prevent dCas9 tethering to DNA. Since these early anti-... AcrIIC2 interactions with Cas9 interferes and prevents the correct assembly of the crRNA-Cas complex ...binds to the HNH endonuclease domain of Cas9 and prevents target DNA cleavage (Harrington et al., 2017)....vectors. Having a way to shut off CRISPR-Cas systems prevents the off-target effects and cytotoxicity that result...

  2. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...explosion of software tools accompanying the CRISPR revolution. (See omictools for even more.) However, none...step of mutating guide sequences in a way that prevents re-cutting but does not disrupt gene expression...CrispyCrunch is how it mutates guide sequences to prevent re-cutting of the sequence inserted by HDR. Although...mutating a single base pair may not be enough to prevent re-binding. Lastly, the sequence inserted by HDR... target regions Design donor RNA Design primers Review and order reagents Perform your wet lab experiment...

  3. Deep Dive: qPCR

    Type
    Blog Post
    ...and I recommend Bustin, 2017 for a more in-depth review.)  Your amplicon should be between 70-200 bp long...ideally located close to either the forward or the reverse primer.  There is a list of useful primer design...efficacy with a standard curve. If relying on previously-generated validation data, be aware that different...transcripts. However, adding too many reagents can prevent your reaction from reaching the plateau stage after...sample compared to your control. But as we said previously, just because it’s used often doesn’t mean it...York 2014Bustin S; Huggett J. qPCR primer design revisited. Biomolecular Detection and Quantification. Volume...

  4. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ...thus structural epitopes, but can reveal linear epitopes previously hidden within the protein. Created...cross-linking can block the epitope of some targets, preventing your primary antibody from binding (Fig.1). The...cleaner results by not permeabilizing as you’ll prevent binding to any target protein that is still on ...preserve sample morphology, while organic solvents can reveal otherwise hidden epitopes. However, keep in mind...

  5. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ... unraveling the mysteries of behavior. We’ve previously discussed some of the general considerations ....  What’s involved in locomotor sensitization? Previous research has shown that regions such as the nucleus...transmission leads to locomotion sensitization. (for review see van Huijstee & Mansvelder  2014). Scientists...the locomotion of the animal. This confirms the previously suggested inhibitory mechanism of locomotor sensitization... times are ahead for behavioral neuroscience to reveal insights into the complex relationship between ...

  6. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    ...experimental treatment can be monitored. This can reveal previously unknown functions for genes, and lays the ...of the zebrafish genome was published in 2013, revealing that about 70% of human genes have a counterpart... ways. Zebrafish mutants In the 1990’s, two revolutionary large-scale random mutagenesis screens produced...unlocked novel research into genetic compensation, a previously unknown mechanism of gene regulation (El-Brolosy...

  7. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...always to the same extent. As described in our previous Origin of Replication post, DNA replication is...bacteria Before looking at an example plasmid, let's review how E. coli normally replicate their chromosomal...sequence forming this pseudoknot, so its binding prevents the mRNA from folding and blocks RepZ translation...high levels of the lncRNA bind to RepZ mRNA and prevent this pseudoknot from folding, blocking replication...bacterial plasmids. Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/...

  8. Data Freedom: The Expansion of Data Sharing in Research Publications

    Type
    Blog Post
    ...publications. The policy was not an overnight revelation, rather it was the result of consultations between... rationale includes increasing the ability of reviewers (and, once published, anyone) to detect possible...apply different or novel tools and assumptions to reveal findings that the originator was oblivious to, ...called Scientific Data, which aims to act as a peer-reviewed repository for “scientifically valuable” datasets...or table, clicking on the published chart could reveal the underlying data and allow other analyses “on...

  9. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...gene are amplified. RPA reactions also contain reverse transcriptase to produce cDNA. Isothermal amplification...are designed, each binding to one strand of the reverse-transcribed target sequence that are present close...with 9V battery Approach :This preprint uses a previously developed method to detect SARS-CoV-2 in the ...samples. First, RNA is extracted and subject to reverse transcription. This is then amplified by PCR using...detect fluorescence upon cleavage. Highlights: Reverse-transcribed RNA samples are amplified by PCR instead...

  10. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...Saccharomyces cerevisiae. Paques F and Haber JE. Microbiology and Molecular Biology Reviews (1999). PubMed...repair and genome engineering CRISPR-Cas9 has revolutionized the genome engineering world and made targeted... disrupting one or both of these sequences. To prevent re-cutting and the resulting mutations after an...transcribes RNA encoding your repair template, then uses reverse transcriptase to generate the complementary ssDNA...

  11. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ...s post and give it a final read Browse list of previous depositors from Boston University Contact Dr. ...that post from Eric before doing anything else. Preventing procrastination Even if you’ve done all the prioritizing...planning in the world, procrastination can easily prevent you from making progress on a task. For instance...to go sit in the tissue culture hood may have prevented me from splitting cells until late in the day ...podcasts. JUST GET STARTED - One Harvard Business Review article on procrastination recommends testing out...

  12. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    ...lack of complete information does not, however, prevent us from identifying problems with the structure...take on opportunities to do some blogging, write reviews, or write for a university publication, improving...was on diversity in academic research. As during previous panels, speakers Moon Duchin, Rafael Luna, Joan... need some way of capturing, quantifying, and preventing this type of failure. Final thoughts The FOR ...Additional Resources The Postdoctoral Experience Revisited. PubMed PMID: 25590106. Paula Stephan’s Book -...

  13. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...efficiency of 30-60%. This is much more favorable than previous methods yielding 1-10% knock-in. Read on to learn...transcription, and use the IVT product as a template for reverse transcription to create your ssDNA. Note that the...conditional deletion of exon 6 of Syt1, mimicking a previously made mouse model. To avoid disrupting splice ...separates the gRNA target sequences from their PAMs, preventing re-cutting after editing. How to use Addgene ...Additional Resources on the Addgene Blog Read Our Previous Post on CRISPR Mouse Models Learn about Homology...

  14. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    ...applications. Chemical Reviews, 121(20), 12384–12444. https://doi.org/10.1021/acs.chemrev.1c00260 Wilson, D...quicker than others. We’ve covered some methods in previous blog posts, like developing recombinant antibodies...antibodies for autoimmunity and inflammation. Nature Reviews Immunology, 10(5), 301–316. https://doi.org/10.1038...for the directed evolution of proteins. Nature Reviews Genetics, 16(7), 379–394. https://doi.org/10.1038...

  15. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ... the University of California, San Diego. The revolution in genetic engineering techniques is a speculation...and just when the Mendelian genetics was being revived by the botanist Hugo de Vries (1896).  In 1928,...Blade Runner (1982). It is a dystopian film that revolves around synthetic humans called replicants which..., and C. In this movie Ethan Hawke’s character revolts against class divide that is created by genetics...prime editor which is a fusion between Cas9 and reverse transcriptase. Using an elongated template + guide...

  16. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...transfer worked with Ponceau S staining or other reversible protein staining immediately after your transfer...expecting low amounts of protein, make sure your reversible staining method of choice is appropriately sensitive...transfer buffers. To slow down smaller proteins and prevent them from over-transferring, increase the amount... away the SDS. To speed up larger proteins and prevent them from under-transferring, try increasing the...Technical errors Search online/ask an expert Review protocol videos for a visual guide to good technique...Check transfer with Ponceau red stain or other reversible stain* Roll out all of the bubbles in the...

  17. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...real time. Two-photon microscopy was the next revolution in cell imaging, giving researchers the ability...(intravital imaging) studies it’s enabled are revealing the cellular behaviors that mediate adaptive and...proteins (protein that undergoes light-induced irreversible photoconversion from green fluorescence to red... 1. Peters, Nathan C., et al. "In vivo imaging reveals an essential role for neutrophils in leishmaniasis.... Egen, Jackson G., et al. "Intravital imaging reveals limited antigen presentation and T cell effector...

  18. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...immune cells harvested from patients and thereby prevent cancer cells from using this receptor to evade ...immunotherapies currently under study. Check out this review for an in-depth look at a variety of up and coming...monoclonal antibody-based therapeutics." Nature Reviews Cancer 15.6 (2015): 361-370. PubMed PMID: 25998715...Lloyd J. Old. "Antibody therapy of cancer." Nature Reviews Cancer 12.4 (2012): 278-287. PubMed PMID: 22437872...Cancer immunotherapy via dendritic cells." Nature Reviews Cancer 12.4 (2012): 265-277. PubMed PMID: 22437871...

  19. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    ...optogenetics. Annual review of neuroscience. 2011;34:389-412. doi: 10.1146/annurev-neuro-061010-113817....effectiveness of your optogenetic manipulations (though a reviewer will probably ask for electrophysiology data too...best suit your specific experiment. Consult these reviews for a discussion of the different variants [1, ...underlying brain disease in animal models. Nature reviews Neuroscience. 2012;13(4):251-66. doi: 10.1038/nrn3171...

  20. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    ...integrate protein tags into the genomes of S. cerevisiae and S. pombe. The protocol is surprisingly simple... in yeast. These tagging vectors are based on previously described pFA6a-link vectors and include a Kan...assessed many of these fluorescent tags in S. cerevisiae, looking at their performance in categories such...a complimentary set of plasmids for use in S. cerevisiae, with the additional benefit of multiple selection...
Showing: 181 - 200 of 741 results