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We narrowed to 777 results for: REP

Showing: 321 - 340 of 777 results

  1. Design Tips for Prime Editing

    Type
    Blog Post
    Published
    Jan. 23, 2025, 2:15 p.m.
    ...that meant rereading many of the original papers reporting various prime editing tools. These papers are ...quick refresher: prime editing is a “search and replace” genome editing technique that relies on a fusion...or longer) tracts of edited bases. DNA mismatch repair (MMR) is less efficient at identifying these “bubbles... use the edited strand as a template during DNA repair. Test multiple nick sites, starting with sites ..., Raguram, A., & Liu, D. R. (2019). Search-and-replace genome editing without double-strand breaks or ...

  2. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ...us! A plasmid is a small piece of DNA that can replicate independently from its host’s chromosome. Although... inherited by daughter cells during cellular reproduction, similar to the host’s chromosome, or they can...For example, helper plasmids were developed to replace helper viruses in AAV production systems. We’re...the many plasmid tools available in the Addgene repository. To learn more about some other common plasmid...plasmid features, such as origins of replication and promoters, check out our other Plasmids 101 posts, and...

  3. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog Post
    Published
    July 12, 2021, 1:15 p.m.
    ...including a handful of plasmids that have been in the repository for many years, has now grown into a collection...neutralization assays, part of a study that examined the repertoire of antibodies from 19 COVID-19 convalescent individuals...tools based on Cas12a and Cas13  which could be repurposed for SARS-CoV-2 RNA detection. Early in the pandemic...above, some of these plasmids have been in the repository for many years while many were developed during...Harrison SC, Wesemann DR (2021) Memory B cell repertoire for recognition of evolving SARS-CoV-2 spike ...

  4. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    Published
    Sept. 26, 2019, 12:50 p.m.
    ...modification with Homology Directed Repair (HDR), CRISPR activation or repression, transgenic mouse line creation... which means they are non-infectious and non-replicative. Nanoblades are produced by transfecting the ...compared to DNA transfection.  Homology directed repair (HDR) with Nanoblades CRISPR-based HDR allows for...transduction efficiency by neutralizing the charge repulsion between virions and the cell surface, and also...

  5. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    Published
    Aug. 11, 2016, 2:23 p.m.
    ... on the composition of the viral vector and the repeat sequences present in the LTR, the vector may undergo...undergo genomic rearrangements during replication. To avoid genomic rearrangements, try amplifying the... and the type of virus, there have been varied reports of titer losses from 5% to 50% per freeze thaw ...exchanging the medium after transfecting cells, replace the medium with half the volume typically used ...virus particle to a target cell by reducing the repulsive electrostatic forces between these two negatively-charged...

  6. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    Published
    July 14, 2016, 2:30 p.m.
    ...are difficult to modify through homology-directed repair. Both Cpf1 and its guide RNAs are smaller than ...in mammalian cells, LbCpf1 and AsCpf1. In both reports, on target editing efficiency for the Cpf1 orthologs...mismatches, but variably so, with Kleinstiver et al. reporting that Cpf1 can tolerate mismatches at gRNA positions... may improve the frequency of homology-directed repair (HDR) over non-homologous endjoining (NHEJ). Cas9...chance” mechanism. To prevent new editing post-HDR, repair templates should remove the PAM sequence. Many ...

  7. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    Published
    Oct. 17, 2017, 12:55 p.m.
    ...eliciting questions like “what does this model representt?" ... "how does this protein function in the ... nuclease active site wasn’t even close to its reported cleavage site on the complementary DNA strand....useful. While a physical model of a protein might represent one aspect of the protein very well, it will almost...is also wrong in at least one serious way. We represented the Cas9 protein in a surface format made of ...model also displays the several Lys/Ala mutations reported by Ian Slaymaker and colleagues to reduce off-...

  8. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

    Type
    Blog Post
    Published
    Dec. 3, 2019, 11:38 a.m.
    ...changed when we changed from searching for viral preps instead of plasmids? The available filtering options...to only show plasmids with ready-to-use viral preparations available Services: If there are viruses available...filter will allow to refine by serotype or lentiviral prep Requests: Only see plasmids with more than the indicated...distributed to industry research scientists Viral prep filter categories Promoters: Select the best promoter...type Serotype: Choose the AAV capsid or lentiviral prep needed Tags: Identify a specific marker protein ...

  9. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    Published
    Aug. 14, 2018, 12:42 p.m.
    ...synthetic peptide FLAG, the synthetic streptavidin-binding strep-tag and, more recently, OLLAS (Escherichia...STORM imaging, and electron microscopy. Sample preparation for these techniques sometimes makes it more ...retain their ability to be detected after sample preparation better than standard proteins tags like GFP. ...Addgene Browse all AAV vectors Find ready-to-use AAV preps ...

  10. How to Write a Scientific Review Article

    Type
    Blog Post
    Published
    Feb. 16, 2017, 3:30 p.m.
    ...this is represented by factorials. In environmental science and ecology, this can be represented by the... member of a class of proteins was mutated, as reported in various studies. We made a table that listed...for each protein we listed all the studies that reported mutations in that protein (including how frequently...areas are worth exploring.  Edit and rewrite, then repeat If you’re like me, your rough draft is really rough...

  11. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    Published
    Nov. 1, 2018, 12:35 p.m.
    ...CcdA (antitoxin) where CcdB inferes with DNA replication by binding the GyrA subunit of DNA gyrase and...Successful cloning occurs when the ccdB gene is replaced by the gene of interest. As a result, cells that...This MazF-ZeoR cassette is flanked by two direct repeats which can be used for marker removal so that the...second recombination event between the two flanking repeats. In cells where recombination has not occured, ...concept, they tested a lux-mediated bioluminescence reporter in vivo in the absence of selective pressure and...

  12. Healthcare Consulting: A Door to the Business of Life Sciences

    Type
    Blog Post
    Published
    Oct. 13, 2016, 2:30 p.m.
    ...personality types, and to devise strategies to be prepared for addressing unexpected situations. At the end...long-term? A few years as a healthcare consultant can prepare you for a successful career in several different...foot in the door When I was first considering preparing for and transitioning to a career in healthcare...learning about the nature of the job and the ways to prepare for it. Besides those conversations, reading online...Michael Koeris' Journey from Grad Student to Entrepeneur Get Tips on Resume Writing Resources on Addgene.org...

  13. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    Published
    March 28, 2024, 1:15 p.m.
    ...and signaling, as well as immune response, tissue repair, and kidney development. Despite their ubiquity...integrin antibodies available through Addgene’s repository. The collection includes 26 unique antibodies...in competition with other similar integrins, as reported by IPI and the Springer lab (Hao et al., 2024)...In this study, we, along with the Springer lab, reported the binding specificity, kinetics, and affinity...reconstructions of the original hybridomas as new reproducible rat or mouse rAbs. The other eight are chimeric...

  14. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    Published
    Dec. 2, 2019, 2 p.m.
    ...the glycoprotein, avidin or a similar protein, streptavidin. This strong interaction allows scientists to...hours biotinylated proteins can be captured on streptavidin affinity beads or matrices. The bait protein... where two proteins are fused to complementary reporter proteins that can only assemble when in close ...Biotin tagged proteins can then be purified using streptavidin and separated via flow cytometry.  Find plasmids... ascorbate peroxidase as a genetically encoded reporter for electron microscopy." Nature biotechnology...

  15. Antibodies 101: Normalization and Loading Controls for Western Blots

    Type
    Blog Post
    Published
    April 8, 2025, 1:15 p.m.
    ...in the same lane, and the resulting fraction is reported. Figure 2: Normalization with loading controls...equally strong, so the normalized value simply reports which POI band is stronger. The right gel has exactly...abundance on your membrane, which would be directly reported by band intensity. Researchers use later steps...perturbed by the experiment Unfortunately, multiple reports suggest that common housekeeping proteins are influenced...lysate (10–40 µg) probed with β-actin (1:2000). Representative of three independent blots. B–C) Graphs showing...

  16. Twenty Years of Sharing Science!

    Type
    Blog Post
    Published
    Jan. 8, 2024, 3:25 p.m.
    ... When we started in 2004, we were the plasmid repository. In 2016, we added viral vectors, and in 2022...ready-to-use antibodies. Now, we think of ourselves as a repository for DNA-based materials, including our popular... kits and libraries. This DNA-based materials repository contains 139,868 reagents from 5,821 labs in ... system for scientists using materials in our repository, from our customer and science support to our...

  17. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    Published
    Oct. 2, 2018, 12:56 p.m.
    ...really like? Gather data of course. Ask questions. Prepare by going sleuthing on the lab website, publication...well. What do they have to say about this lab’s reputation? If you are choosing a training lab, I strongly... meaningless tasks, removal of responsibility, repeated reminders of blunders, setting target up to fail...ensure that you are expanding your options and preparing for both academic and non-academic career paths...most papers Productive external collaborations Reputation for not being open PI helps lab members determine...

  18. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    Published
    Sept. 29, 2016, 2:30 p.m.
    ...to their utility in the health sector, phages represent a reservoir of carbohydrate binding proteins waiting...successfully identified using this method. Step 1: Prepare a random fragment library of phage DNA This first...with cloning large fragments. After fragmentation, repair sheared ends with T4 DNA polymerase (to create ...purification, but overnight at 30°C is a good start. Prepare a lysis solution containing your favourite bacterial...20°C (or -80°C) followed by 5 min at 37°C (each repeated 5 times) works well. Leaving the membrane overnight...

  19. Building Global Connections with the International Mentorship Program USA-EUROPE

    Type
    Blog Post
    Published
    Nov. 6, 2015, 3:30 p.m.
    ...graduate studies. Students also need more access to reputable, organized networks connecting international experts...the University of the Basque Country. Mentees represented a diverse group of individuals with technical...writing fellowships, giving oral presentations, and preparing for job interviews, expanded profesional networks...Spanish scientific community overseas. Official representatives from different Spanish universities and high...development of knowledge, experience, resources, and entrepreneurial visions thereby opening new frontiers for young...

  20. Addgene Protocols Go Viral: AAV Purification and Titration

    Type
    Blog Post
    Published
    June 13, 2019, 12:19 p.m.
    ...in a given AAV preparation. For detailed instructions and calculations on how to prepare a plasmid stock...Addgene has shipped thousands of AAV and lentiviral preps to scientists around the world. In the years since...improve both the yield and quality of the viral preps. For example, last year we published an article ...
Showing: 321 - 340 of 777 results