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We narrowed to 481 results for: vent

Showing: 121 - 140 of 481 results

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

    Type
    Blog Post
    ...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...

  2. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ... cancer. That’s exactly what the lab of Andrea Ventura at the Memorial Sloan Kettering Cancer Center did...delivery of CRISPRS/Cas9 using the Gonçalves and Ventura labs’ plasmids, including protocols, check out ...Crippa A, Rekhtman N, de Stanchina E, Lowe SW, Ventura A (2014) In vivo engineering of oncogenic chromosomal...

  3. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...correctly recombined cells. Recombination is a rare event, so you must select as opposed to screen for cells...participating in development or other physiopathological events to which the chosen promoter responds. You can ...occur in multiple ways. The desired recombination event will remove NeoR only and leave exon 2 floxed, as...

  4. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    ...want the works: organizing reagent storage and inventory, managing orders, tracking students and research...access your protocols at the bench, keep your inventory at your finger tips, and make shopping lists as...protcols, sign-up for shared equipment, and track inventory and orders. Quartzy caters to life science labs...

  6. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...asking the cell to perform multiple recombination events, all linear fragments need to get into the same...cloning requiring just one or two recombination events, homemade competent cells of 107 CFU/μg are perfectly...multiple modifications requires multiple recombination events to assemble the final plasmid. At least 5 fragments...

  7. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...to study the inner workings of the synapse by conventional light or fluorescence microscopy (Dani, et al...allowing you to resolve individual points with a conventional, diffraction-limited light microscope.  Along.... But many ExM protocols are compatible with conventional antibodies, chemical dyes, and genetically encoded...

  8. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ...within a CRISPR guide can prevent the guide from binding to the genome or prevent Cas9 from cutting, both...Perturbation Platform to modify the CRISpick algorithm to prevent guides from being designed that target regions ...

  9. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...editor invented. ABE Adenine Base Editor Converts A → G; the second type of base editor invented. CGBE... CBE; the first type of base transition editor invented.   Abasic site A DNA site with the nucleobase...

  10. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...strike a balance: replicating too slowly will eventually cause a plasmid to be lost from the population...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...

  11. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...Vectors – inversion only once All recombination events mediated by FLP or Cre are reversible. Whereas ... within that sequence and should therefore be prevented. The loxP and FRT target sites have been engineered...Flip-excision vectors) that only allow one inversion event when it was discovered that it is not the exact ...

  12. Lab to Office Culture Shock

    Type
    Blog Post
    ... I talked to a graduate student at a networking event recently who referred to her lab as her family three...career. Yet even success in academia means you’ll eventually end up as a PI working what is essentially an...the move from lab to office will likely happen eventually. If you approach that transition dreading the...

  13. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...propagated better than plasmids without this DNA. They eventually designated this fragment as the ccdB region (...removed the cassette through a second recombination event between the two flanking repeats. In cells where...groups to bases at specific nucleotide sequences, preventing the cognate restriction enzyme from cleaving ...

  14. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ...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 ...

  15. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...sequential homologous recombination events, but because these events are rare, each requires selection....co-opted toward that aim (i.e. sacB). Importantly, to prevent activation of toxins before the second crossover...

  16. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...unlikely to be found in your gene of interest) and prevent incorporation of extensive sequences used for cloning...Translation of the standardized leader peptide prevents secondary structures in the mRNA encoding your...genes from a single transcript, or if you want to prevent read-thru transcription from resulting in the production...

  17. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    ...Dr. Daniel Gibson and colleagues at the J. Craig Venter Institute in 2009. Here at Addgene we added this...chromosome fragments. CRISPR cutting was used to circumvent the need to PCR amplify the fragments of the ... References: 1. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA 3rd, Smith HO. Enzymatic assembly...

  18. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    ...of the symposium and live tweeted throughout the event (look for a ton of tweets from the Symposium under...lack of complete information does not, however, prevent us from identifying problems with the structure... need some way of capturing, quantifying, and preventing this type of failure. Final thoughts The FOR ...

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

    Type
    Blog Post
    ...or point mutations, the 8-10 month process of conventional gene targeting is lowered to about 2-3 months...NHEJ-repaired indel, often occur. Figure 1: In conventional gene targeting (A), you must first generate ...separates the gRNA target sequences from their PAMs, preventing re-cutting after editing. How to use Addgene ...

  20. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ...editor invented. ABE Adenine Base Editor Converts A → G; the second type of base editor invented. CGBE...bound, keeping the two DNA strands separate and preventing repair based on the complementary strand. This...
Showing: 121 - 140 of 481 results