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We narrowed to 956 results for: REP

Showing: 361 - 380 of 956 results

  1. Lab Automation at Addgene

    Type
    Blog Post
    ... The FXP id also able to do high throughput DNA preps and other molecular work if we need it to. This ...Automating DNA purification We're developing ways to prep large quantities of DNA because some countries just...bacterial samples at the same time as opposed to preparing each sample individually. Enter the BIOROBOT! ...BIOROBOT Universal Platform and it suits our plasmid prepping needs perfectly. Working with Qiagen, we developed... the similar buffers as would be used for a mini prep, except it does it on a larger scale. Using the ...between samples. With this automated protocol, we can prep two 96 well plates in just under 3 hours or one ...You might not be doing enough transformations, minipreps, or filling enough vials to automate these processes...

  2. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ...host genome, the virus is replication incompetent, meaning it can’t replicate and continue the infection... tools, thought was given on how to keep the replication of virus and its infectivity in check. Engineering...envelope plasmid), and the polymerase (Pol) for replication and Gag structural protein on yet a third plasmid... this safety feature prevents the virus from replicating and spreading in non-target cells, including ....    Viral cargo-specific safety   After replication competency, the second concern with virus safety...If administering virus to mice, part of your preparation should be doing your best to be well-rested and...familiarize yourself with your organization’s reporting mechanisms before you start an experiment, just...

  3. DIY DNA Ladders from Penn State University

    Type
    Blog Post
    ...State Ladder: Homemade Plasmid Prep Penn State Ladder: Commercial Plasmid Prep Kit Amount 1000 Lanes 1000...significantly cheaper. When using a homemade plasmid prep protocol, it cost less than $10 to make 1000 lanes...bp or the 1 kb ladder. Using a commercial plasmid prep kit increases the costs to $20-30 per ladder: still...DNA ladder? You can find the Tan Lab’s DNA ladder prep protocol here and the pPSU1 and the pPSU2 plasmids...Generating DNA Molecular Weight Markers. Scientific Reports,7(1). doi:10.1038/s41598-017-02693-1. PubMed PMID...Addgene.org Use the Kit-Free Alkaline Lysis Plasmid Miniprep Protocol Learn How to Analyze Your Restriction...

  4. Lab Safety for Biosafety Levels One and Two

    Type
    Protocol
    ...been designed by the Addgene nonprofit plasmid repository and is being shared outside of Addgene for informational...informational purposes only. If you choose to reuse or repurpose this SOP in another location, please note that...

  5. Transferable Skills Guide: Managing a Team

    Type
    Blog Post
    ...unique skills and strengths (you will likely have reports with different levels of experience), find out ...tasks before I was a manager! Second, my direct report was not engaged because I only left him/her with...you’re delegating a recurring task to a seasoned report, there should be questions! Remember that delegating...itself is counter-productive. I always assume that reports want to succeed at work and providing feedback ...feedback is needed for that. In many cases reports are not aware of issues until you bring them up. I wouldn’...wouldn’t go as far as saying that your reports will be grateful for the feedback, but at least you can start...someone’s hard work and reward a job well done. Your report will (needs to) feel valued. Talk to your manager...

  6. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...resulting double strand break (DSB) stimulates DNA repair mechanisms that can be exploited to either inactivate...making targeted modifications to DNA, Cas9 can be reprogrammed to serve as a regulator of gene expression. ...domains such as transcriptional activators or repressors. These fusion proteins retain the ability to ...conventionally used SpCas9 (derived from the bacterium Streptococcus pyogenes). Consequently, fewer genetic tools...observed at the target site.   Figure 3: Gene repression with nuclease-competent Cas9. When complexed ...gRNAs targeting different sites of interest thus represents a straightforward means of exerting tight control...simultaneous cutting and transcriptional activation or repression at the respective sites.   Many thanks to our...

  7. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    ...years at Addgene, we are sharing stories of our repository’s positive impact on science and scientists around...They did this by depositing them in Addgene’s repository. Other researchers, eager to try this new technology...their own to share. Over the years, Addgene’s repository has grown to include many CRISPR tools.  Distributing...Addgene had almost 18,000 CRISPR plasmids in our repository, and we have shared almost 300,000 CRISPR plasmids...of libraries, once made available through the repository. And the number of plasmids and libraries deposited...categorized by year of deposit. Outlines show a representative proportion of CRISPR plasmids requested, categorized...posts are also frequently read: Homology Directed Repair has been viewed 219,289 times, while Validating...

  8. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...species. Reproductive confinement involves working with laboratory organisms that can't reproduce with wild...wild-type chromosome and its own gene is used as a repair template, thereby converting the heterozygote into...experiments are less hazardous in organisms that seldom reproduce sexually because the drive must be much more efficient...confinement methods. Since molecular, ecological, and reproductive confinement typically require very little effort...an accidental gene drive release, which would represent proof positive that at least some scientists could...confinement are those of the author and do not represent a formal stance taken by Addgene or its staff....Insect Populations: I. Cage Studies of Chromosome Replacement by Compound Autosomes in Drosophila melanogaster...

  9. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ...group dependencies  Take, for instance, a recent preprint released by the Boehm and Beroukhim labs at the...research uses ancestry groups rather than self-reported race, it highlights the myriad issues researchers...the results. But the reference genome does not represent the diversity of genetic variation across the ...theory, if the DepMap analysis had an equal representation of every ancestry group, they should have found...designed a tool that could analyze CRISPR sgRNAs and report the likelihood of any given sgRNA having an ancestral... researchers to ignore or miss. The lack of representation in commonly used cell lines led to an incorrect...complicating the issue is the long and ugly history of reporting false essentialist links between biology and ancestry...

  10. CRISPR Antimicrobials

    Type
    Blog Post
    ...containing a phage origin of replication that can be packaged into replication-incompetent phage particles...antibiotic resistance is upon us, and the world is unprepared. Each year in the United States, two million ...essential processes: cell wall synthesis, DNA replication, or protein synthesis. Because they target such...sequences (red). Treating a mixed population, represented here by cells of different colors, specifically...small pools of antibiotic resistant cells could repopulate the environment after treatment. The phagemid...mammalian viral vectors in that the vectors are replication-deficient. Thus, a very large number of phagemids...CRISPR antimicrobials combines them with both replication-competent phages and antibiotics. Phage therapy...

  11. What's New in CRISPR - May 2019

    Type
    Blog Post
    ...transcriptional repressors, they found that fusing dCas9 to an Mxi1 repressor domain repressed transcription... Covalent tethering of DNA repair template to Cas9 Homology-directed repair (HDR) can be used to make ...Gordon's lab developed a strategy to enhance HDR repair using a single-stranded oligodeoxynucleotide HDR...interference (CRIPSRi) platform for targeted gene repression in Candida albicans was developed by Rebecca ...

  12. Making Career Development an Integral Part of the Science Training System

    Type
    Blog Post
    ...Career Development; a Stakeholder Workshop Report.” The report enabled the development of priorities for...Development Hub Workshop Attendees and their 2021 report. Image from pd|hub.   To begin to tackle these...Steering Committee and working groups with representatives from all areas of the scientific training and...the findings were captured in this just released report “Enhancing Dissemination of Evidence-Based Models...programs.  Please join us for the pd|hub Workshop Report Public Release Event scheduled for March 4th from...2:30-4:00 pm ET.  You can learn more about the report's recommendations and join with us to discuss and...

  13. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    ...facilitate repair. Since the development of CRISPR-mediated base editing (BE), these types of repairs can now...the resultant U-G pair is either repaired by cellular mismatch repair machinery making an original C-G...possible through exploiting the homology-directed repair (HDR) pathway which occurs in cells at low frequencies...or reverted to the original C-G by base excision repair mediated by uracil glycosylase (Figure 1). The ...pair (Figure 1). As the cellular inosine excision repair is not as active as uracil excision, ABE does not...al. constructed a number of plant base editors replacing the SpCas9 with the mutant versions of SpCas9 ...

  14. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...experiments. If Cas7-11 lacked this toxicity, it would represent a major step forward for CRISPR-based RNA editing.... Specifically, efficacy rates of Cas7-11 were reported as up to 85%, broadly similar to shRNA and Cas13...However, in those same comparisons, the AbuGoot lab reported no detectable effect on cell viability – meaning...mutations wouldn’t matter – the mutated copy could be replaced by simply dragging and dropping in a functional...allowing a one-size-fits-all approach to gene replacement therapy for most patients. But how does one go...work. (It was also found to be independent of gene repair.) This lowers the chance of the insert being “dropped...techniques like HITI or HDR, which require specific repair pathways to work, PASTE is more specific and efficient...

  15. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...the new plasmids, antibodies, and viral preps in our repository through our Hot Plasmids articles.  Here's...facilitate SARS-CoV-2 research New viral vector preps of Voltron2 voltage sensors AAV vectors for targeting... tRNAs, within the lentiviral 3’ long terminal repeats (LTR). The 3’ LTR is duplicated during lentiviral...antibodies were recently added to the Addgene repository of tools for SAR-CoV-2 research. Two new antibodies...voltage sensors By Brian O’Neill New viral vector preps expressing the Voltron2 voltage indicator, created...-microglia cells.      Figure 5: A) Representative confocal images showing GFP expression from...Disease, including plasmids, antibodies, viral vector preps, CRISPR tools, and more.  ...

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

    Type
    Blog Post
    ...and placed in the legend. Widely reported difficulties in reproducing experimental data are likely, at...readers may locate data supporting the research reported in PLOS publications. The policy was not an overnight...author. Instead, selected images are said to be representative or the statistical variation is calculated ...datasets are too large to transfer to a journal repository, they are likely too large to be retrieved by...Scientific Data, which aims to act as a peer-reviewed repository for “scientifically valuable” datasets. It likely...likely won’t be long before other data repositories emerge that allow efficient searching and extraction... in our interest to ensure that our data are reproducible and useful to others and efforts to enhance ...

  17. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ...painstakingly assessed to something a bit more representative of the communities’ opinion. The trouble is... The foremost handles the design of sgRNA and represents the lion’s share of the currently available tools...a good example is CRISPR-GA. These tools assess repair events by type (NHEJ/HDR) and track indels at a...no longer available), is a simple catalogue of reported sgRNA designs. The second database, CrisprGE (...speed up the sgRNA design process by removing repetitive tasks and/or by providing features to help with...it takes authors to add a genome to the tool’s repertoire. Organism agnostic tools are the better solution...to learn the language and what the parameters represent. The best learn-by-doing tool is E-CRISP. The ...

  18. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...resistance cassette, and the R6K origin of replication to allow replication only in hosts carrying the bacteriophage...mutated allele introduced from a conjugative plasmid replaces the chromosomal copy. This is a versatile technique...recipient of interest. Because pDS132 cannot be replicated in the recipient, the only antibiotic resistant...modifications to pDS132 (Philippe et al., 2004): Replaced the sacB cassette with one of three toxins from...the Salis group) (Espah Borujeni et al., 2014). Replaced the pDS132 MCS with a versatile, synthetic MCS...recombineering enables simplified chromosomal gene replacement in Escherichia coli." Appl. Environ. Microbiol...genome editing in Escherichia coli." Scientific reports 5 (2015): 15096. PubMed PMID: 28060411. Thomason...

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

    Type
    Blog Post
    ... are tagged with streptavidin, the lateral flow strip first captures the streptavidin portion of the probe...DNA polymerase, recombinase, and the ssDNA-FQ reporters that exhibit fluorescence upon cleavage. Upon ... results in the trans cleavage of the ssDNA-FQ reporter resulting in fluorescence. Cas12a binds all the...results in the continuous cleavage of the ssDNA-FQ reporter. The resulting fluorescence can be observed under...fluorescence visualizer with 9V battery Approach :This preprint uses a previously developed method to detect SARS-CoV...biotinylated primers. These are then immobilized with streptavidin coated beads. Fluorescently labelled dCas9 complexes... concept to lateral flow strips that contain streptavidin which can bind to the biotinylated targets. ...

  20. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ...molecular brightness (Table 1).  NIR reporters and biosensors NIR reporters can tell us about cellular events...higher apparent cellular brightness than previously reported widely used dimeric iRFPs (Table 1). In general...protein-protein interactions, we recommend using NIR split reporters based on miRFPs (Shcherbakova et al., 2016), since...fluorescence of co-transfected EGFP. e Originally reported as a monomer16, IFP2.0 was later found to be a...E., Sivanesan, D. & Michnick, S.W. An infrared reporter to detect spatiotemporal dynamics of protein-protein...Filonov, G.S. & Verkhusha, V.V. A near-infrared BiFC reporter for in vivo imaging of protein-protein interactions...evolution approach applied to fluorescent voltage reporters. Nat Chem Biol 14, 352-360 (2018). Yu, D. et al...
Showing: 361 - 380 of 956 results