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We narrowed to 79 results for: plasmid cloning by pcr

Showing: 71 - 79 of 79 results

  1. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...encoded on separate plasmids and are co-transfected into cells alongside a plasmid encoding the minigenome...four viral genes in the same plasmid (as opposed to four separate plasmids), there are a few reasons arguing...each would be relatively large. Cloning the viral genes in this plasmid would be more difficult compared...separate (and thus smaller) plasmids. Finally, having four separate plasmids allows researchers to partially... 2 μg minigenome plasmid per well (in a 6 well plate format). However, since plasmid quality and transfection...Minigenome System). From a more technical standpoint, a plasmid with all four genes and necessary control elements...detected directly by Northern blot analysis or RT-PCR. Uses of the minigenome system Minigenome systems...

  2. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    Published
    Nov. 21, 2024, 10:47 p.m.
    ... that made it easy to share plasmids, both for people depositing plasmids and for people requesting them... Collections Lentiviral Plasmids Tetracyline Inducible Plasmids Empty Backbones Bacterial Expression...Expression Systems AAV Plasmids Blogs Origin of Replication What is a Plasmid? How to Design Your... Verify Your Plasmid Using a Restriction Digest Analysis Videos What is a Plasmid? How to Design...video, however, is “What is a Plasmid?”, An animated explainer of a plasmid and its parts that helps the...them. Prior to this, many plasmids were available, but they often weren’t easily accessible, due to a ...barriers, allowing people to access a wide catalog of plasmids from all over the world. This idea, that available...

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

    Type
    Blog Post
    Published
    April 5, 2018, 12:42 p.m.
    ...editing. How to use Addgene plasmids for Easi-CRISPR You can use Addgene plasmids to create a repair template...of Addgene plasmid options for templates - see the example at right using blue flame plasmid pmScarlet_...Easi-CRISPR repair templates using Addgene plasmids. Design PCR primers to add T7 promoter-left homology...desired insert (e.g., mScarlet).   Plasmid ID Plasmid Name Tags 69090 pMJ915 MBP (N ...out our CRISPR Guide Find Plasmids for Cas9 Purification Find CRISPR Plasmids for Mammalian Expression ...pre-CRISPR was very arduous. First, you’d construct a plasmid with your desired changes, long homology arms and...mouse embryonic stem cells. After selecting for clones with the recombined product, you’d microinject ...

  4. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...protein When expression plasmids are available, or the necessary expertise to clone them, it can be useful... includes clone names, which can allow you to search different providers for the same clone. This can ...assessment of labeling pattern between multiple antibody clones targeting the same protein (independent antibodies...involves the transient transfection of expression plasmids in a given cell line. For flow cytometry, it is...protein of interest. HLDA workshop approved antibody clones The antibody validation process represents a high...For CD365, they examined two different antibody clones produced by different vendors. The specific epitope...where different manufacturers may have the same clone available with different conjugates. Summary Flow...

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

    Type
    Blog Post
    Published
    July 12, 2021, 1:15 p.m.
    ...received over 13,000 plasmids requests for COVID-19 related plasmids. These plasmids have reached labs in...small collection of plasmids for coronavirus research, including a handful of plasmids that have been in...some highlights of the plasmids shared over the last year and how these plasmids have been used to accelerate...circulating and plasmids containing these mutations have been deposited at Addgene. Plasmids deposited from... labs also deposited plasmids for producing COVID-19 diagnostic enzymes. Plasmids in this toolkit include... deposited plasmids expressed 26 of the 29 SARS-CoV-2 proteins which all but one are cloned into pLVX ...marker for stable cell clone selection. For more information on these plasmids and a behind-the-scenes...

  6. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    Published
    March 15, 2016, 2:30 p.m.
    ...quantify the amount of PCR product that comes from amplification of the original plasmid which can over-inflate...for expression of a gene carried on the transfer plasmid or quantifying the number of viral copies that ...beneficial as they ensure that the correct transfer plasmid was used in the transfection; if preparing several...antibiotic resistance genes carried on transfer plasmids. Target cells are transduced with serial dilutions... titer are direct p24 measurements via ELISA and qPCR for viral RNA. Measuring p24 levels in a viral prep...first converted to cDNA and then quantified using qPCR primers targeting specific viral components such...assays depending on the vector backbone. When using qPCR based methods for viral titration consider using...

  7. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ...transgene   Transfer plasmids The transfer plasmid is the vector you clone your gene of interest into...virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing all the structural genes...crossover event between the vector plasmid and the packaging plasmid caused the Psi sequence to be placed...usually 293T cells, with your transfer plasmid and your packaging plasmids. For this, you can use the transfection...techniques are still used and efficiently transfer plasmid DNA efficiently into dividing immortalized cells... (Gag, Pol, Tat, Rev and Env) and the transfer plasmid encoding the insert of interest as well as sequences...the packaging signal Psi). Because the packaging plasmids lacked the psi signal, the genes encoding the ...

  8. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...They generated plasmids tagged with EB3 to mark the growing tips of microtubuli and plasmids tagged with ...and dCas9-VPR targeting plasmids can be found at Addgene. Check out these plasmids for piggyBac gRNA delivery...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head... Find the plasmids at Addgene The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll...

  9. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    Published
    July 13, 2023, 1:15 p.m.
    ...single plasmid transformation, though, a pooled library has hundreds to millions of distinct plasmids, all...anyone who has worked with individual plasmids knows, some plasmids amplify well and others don’t. The size...pooled library, which contains a collection of plasmids in a single tube, or a pre-made viral prep library... prepare the libraries for you.  Starting from plasmids requires amplification, verification, and lentiviral...library and appropriate packaging and envelope plasmids into HEK cells allows production of lentivirus...amplify the library. This is similar to amplifying a plasmid: transform E. coli with the pooled library DNA,...own protocol that amplifies without disrupting plasmid representation. A good place to start is with a...
Showing: 71 - 79 of 79 results