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We narrowed to 956 results for: REP

Showing: 441 - 460 of 956 results

  1. DNA Purification Without a Kit

    Type
    Blog Post
    ...purification kits. Kit-free alkaline lysis plasmid miniprep Starting material: 2 mL bacterial culture Product...Product: plasmid DNA This kit-free plasmid miniprep protocol from Addgene follows a similar workflow as ...authors estimate that a gravity-based column plasmid maxiprep kit takes 1-1.5 hours to complete, while this ... minutes. Additionally, while commercial plasmid prep kits come in discreet sizes (mini, midi, maxi, giga....0007750  Vogelstein, B., & Gillespie, D.A. (1979). Preparative and analytical purification of DNA from agarose...Purification and Gel Extraction Kits. Scientific Reports. https://doi.org/10.1038/s41598-018-30316-w  Zou...

  2. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...transformation   Spectinomycin/streptomycin Spectinomycin and streptomycin both inhibit protein synthesis...expensive antibiotic compared to streptomycin. Spectinomycin/streptomycin can be successfully used for plant...offers is marginal. Fortunately, amp can easily be replaced with carb if satellite colonies are an issue. ...spectinomycin). Cons: Cost (spectinomycin) or stability (streptomycin), doesn’t work for all bacteria.   Zeocin  Zeocin...

  3. An Introduction to Adenovirus

    Type
    Blog Post
    ...transcription) is demarcated by viral genome replication (Harrach, 2014). Given that the early genes ...encapsidate new virions) are not expressed. Note that replication-competent AdV vectors also exist and are used...the final topic, which is that natural AdVs, replication-competent viruses with a recombinant genome, ...(Tollefson et al., 1996) – cell death has been reported when using vectors, and immune inactivation of...pre-existing toolbox for adenoviral vectors in Addgene’s repository is smaller than for other vector types, we hope...Hermiston, T. W. (2003). Multigene expression from a replicating adenovirus using native viral promoters. Mol ...

  4. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...nuclease-deactivated Cas9 (dCas9) can be tethered to a repressor (CRISPRi) or an activation domain (CRISPRa) to...CRISPRko sgRNA library as pooled library or lentiviral prep! Dolcetto: human CRISPRi sgRNA library While CRISPRko... protein (dCas9). dCas9 is then tethered to a repressive domain, such as KRAB, to prevent efficient transcription...Dolcetto library! Now also available as a lentiviral prep. Calabrese: human CRISPRa sgRNA library In addition...Find the Calabrese pooled library and lentiviral prep here! Applications for the Root and Doench CRISPR..."Genome-scale CRISPR-mediated control of gene repression and activation." Cell 159.3 (2014): 647-661. ...

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

    Type
    Blog Post
    ... Institute) has consistently been one of the repository’s most frequently requested plasmids. I wondered... and you decide to share them using Addgene’s repository, you can also share your associated protocols...the software aims to be a life science protocol repository. The software can be used with the ZappyLabs ... protocols listed on OWW, including those for preparing your own competent cells. If you’re familiar with...Protocol Exchange (NPE) is an open online protocol repository hosted by the Nature Publishing Group. Like the...protocols when they deposit their plasmids to the repository. These protocols show up on the related plasmid...

  6. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ... in cancer studies, act as a reporter for DNA double strand break repair, and to label pathogenic intracellular...maintaining fluorescence. The protein structure, first reported in 1996, is an eleven β-sheet-containing “barrel...been done to expand the red-fluorescent protein repertoire; however, these proteins are unique from GFP ...constructing fluorescent fusions. Transcription Reporter: Placing GFP under the control of a promoter of...a given cell type. This type of transcription reporting was among the earliest uses of GFP. Fluorescence...

  7. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

    Type
    Blog Post
    ...in our repository that you can use to study these genes! Groucho, a transcriptional co-repressor The first...after the protein Groucho, a transcriptional co-repressor important in development. Groucho family proteins...specific set of DNA-binding transcription factors to repress the expression of genes primarily involved in development... see for yourself. We have one plasmid in our repository, plasmid 39624, that contains cmr.   tinman and... Groucho/TLE/Grg family of transcriptional co-repressors." Genome biology 9.1 (2008): 205. PubMed PMID...

  8. Hot Plasmids - November 2023

    Type
    Blog Post
    ...the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. Today... set of fusogenic and targeting components to reprogram delivery vehicles like lentiviral vectors, CreVLPs...doi: 10.1038/s41592-023-01880-5. PMID: 37188954. Preprint available on bioRxiv – doi: 10.1101/2023.03.09.531939...doi: 10.1016/j.cell.2023.05.044. PMID: 37385249. Preprint available on bioRxiv – doi: 10.1101/2023.02.07.527548...doi: 10.1038/s41587-022-01524-7. PMID: 36593408. Preprint on bioRxiv — doi: 10.1101/2021.10.13.463862.  ...

  9. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ... strand break (DSB) mediated homology dependent repair by enhanced green fluorescent protein (EGFP) reconstitution...sgRNA guided Cas9 endonuclease, homology dependent repair (HR, homologous recombination; SSA, single strand...halves of EGFP to recombine by homology directed repair, and resulting in the expression of EGFP. Using...Samantha Young is a Medical Writer with a PhD in reproductive biology and genetics. She loves to communicate...expressing Cas9 and single guided RNA." Scientific reports 3 (2013). PubMed PMID: 24284873. PubMed Central...Non Homologous End Joining and Homology Directed Repair Additional Resources on Addgene.org Find Software...

  10. PEI Calculator for Planning AAV Packaging Transfections

    Type
    Blog Post
    ...Transfer Plasmid : 1 pHelper: 1 RepCap OR 3 Transfer Plasmid : 5 pHelper: 2 RepCap Select what type of tissue...goes, “failing to plan is planning to fail.” Preparation is a key step to any experiment and can help ...experimental details on the spreadsheet: Name your viral prep Enter the specific names of the plasmids you plan...using 28 μg of total plasmid (Transfer, helper, and RepCap added together) per 15 cm dish or 2500 μg of plasmids...

  11. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...similar protein that was discovered in the 1960s, strepavidin. The Biotin-avidin complex can then be used and...primary antibody respectively. Avidin bound to a reporter is then added to the reaction for easy detection... experiments. During detection, a biotinylated reporter enzyme that is pre-incubated with free avidin ...protein of interest leading to amplification of the reporter signal and easier detection. This strategy is ...Forster, A. C., Skingle, D. C., & Symons, R. H. “Preparation and uses of photobiotin.” Methods in Enzymology...mammalian cells with biotin ligase and monovalent streptavidin.” Nature Protocols 3.3 (2008): 534–545. PubMed...

  12. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...dCas9-VP64) or repressors (dCas9-KRAB or dCas9-SRDX) can also be used to activate or repress target genes...engineering CRISPR can be used to knockout, activate or repress target genes in plants using the same general experimental...containing plasmids for knockout, activation and repression of target genes in plants and many of the aforementioned...in human cells. In contrast to homology directed repair, these methods do not involve double strand break... the Patron Lab.  As with all plasmids in the repository, we highly recommend reading the associated publications...

  13. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...regulatory elements (to mutate promoters/enhancers in reporter constructs), in functional analysis of proteins...introduced to endogeneous DNA through homology-directed repair (HDR) of a CRISPR/Cas9 induced double-stranded ...to the endogenous target, yet to facilitate the repair, the template should be resistant to Cas9 cleavage...your primers should be free of palindromic and repetitive sequences, but if present, a minor extension ...product (see figure). Template A high purity plasmid prep significantly increases the success rate of site...’ll want to steer clear of these strains when preparing the plasmid template for site directed mutagenesis...

  14. Evolution of Lab Techniques

    Type
    Blog Post
    ...versions obsolete, the techniques we use in lab are replaced by improved versions that save both time and money...observed. Today Southern blotting has been largely replaced by real-time PCR to answer the same experimental...researcher (or a simple computer program) can then reproduce the DNA sequence (Figure 2). Accuracy and accessibility...this technology, radiolabelled nucleotides were replaced with fluorescently labeled ones allowing for fluorometric-based...discoveries (5, 8). While newer techniques have largely replaced these, these techniques still have useful applications...already commercially available although they are reported to have high error rates (10). It is projected...

  15. New Viral Vectors - March 2024

    Type
    Blog Post
    ... our viral vectors repository this year (and it's only March!)  In our repository, you'll find the latest...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share the 21...New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes ...check out Addgene's Data Hub to view viral vector reports from other users. And don't forgot to use the link...

  16. Make a Splash: Notions of Scientific Impact Are Evolving

    Type
    Blog Post
    ...research" (2013) Scientific Reports 3, Article number: 1649 doi:10.1038/srep01649    ... to all articles, Anurag Acharya and colleagues report, the percentage of citations to articles in non-elite...publications. (As an interesting aside, Nature recently reported that the most cited work in history is a 1951 ... the best way to measure impact? As that Nature report also noted, papers detailing Nobel Prize-winning...

  17. Increasing Your Work's Visibility with Addgene: Citation, Search, and Collections

    Type
    Blog Post
    ...plasmid" section Anytime you use a plasmid from the repository, we request that you cite it using both its Addgene...find the fruits of your labor and won’t have to repeat the modification. This will save both time and ...you to browse through the many plasmids in the repository, our scientists have curated a number of special...attention to the great variety of plasmids in the repository. And last, but certainly not least, we often ...Special Collections Pages Suggest Plasmids for the Repository Find Plasmids for Your Research  ...

  18. Antibodies 101: Validation

    Type
    Blog Post
    ...before using an antibody.  Antibodies and Reproducibility First, let’s review why validation is so important...called out antibodies as major culprits in the reproducibility crisis of biomedical research (Baker, 2015;...proposed 5 Pillars of antibody validation which represent five experimental strategies researchers can use...benefit and we can all help improve antibody reproducibility!     References and Resources Additional ...plasmids here!  References Baker M (2015) Reproducibility crisis: Blame it on the antibodies. Nature ...10.1038/521274a  Bradbury A, Plückthun A (2015) Reproducibility: Standardize antibodies used in research. Nature...

  19. Sensing Neuronal Dopamine

    Type
    Blog Post
    ...behaving mice to gather the following types of data: Report time-course and concentration of released DA in...optogenetically activated and inhibited neurons in live mice Report modulation of DA signaling during learning and ...moving mice to gather the following types of data: Report endogenous DA release from a single dopaminergic...looming stimulus in vivo in transgenic zebrafish Report DA release in live mice over the course of Pavlovian...of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...

  20. Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process

    Type
    Blog Post
    ...of a plasmid, the first step is always to send prepped plasmid DNA for next generation sequencing. We ...promoters, selectable markers, tags, origins of replication, and fluorescent proteins that are already included...sequence by NGS. These can include GC-rich and repetitive regions. These regions require additional verification... plasmid off for Sanger sequencing.     If a repetitive region is present in the plasmid, we might need...analyze sequencing results for plasmids in the repository at a glance. Our Snapgene-powered plasmid maps...
Showing: 441 - 460 of 956 results