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We narrowed to 145 results for: transfection

Showing: 41 - 60 of 145 results

  2. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...sequencing to confirm library representation. Transfection of the DNA pooled library and appropriate packaging...Addgene.) This small aliquot is not enough to transfect producer cells, so your first step will be to ...in Lenti-X 293T cells using a polyethyenimine transfection protocol. I also recommend searching PubMed ...

  3. What the HEK?

    Type
    Blog Post
    ...replication. To generate a virus, you need only transfect the ‘helper’ plasmids (viral packaging machinery...Protein production  HEKs are extremely easy to transfect in bulk and produce recombinant proteins at high...

  4. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...mammalian selection cassette to screen for positive transfected cells. The addition of an in-frame fluorescent...you can get away with generating transiently transfected cells, you will not need a mammalian selection...

  5. Antibody Production

    Type
    Collection
    ...expression vectors, human embryonic kidney cells are transfected with the plasmid(s) encoding the heavy and light...contain low levels of co-purified plasmid from transfection. This residual plasmid is extracted and amplified... The fluorescence pattern is compared to an untransfected control. Immunohistochemistry (IHC) Tissue sections...antibody. Protein expression is compared to an untransfected control and the protein size is confirmed. Residual...

  6. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...constitutively expressed to identify successfully transfected cells. C-to-T editing by the APOBEC-Cas9 editosome... to cells segregated solely by a reporter of transfection, which only report the efficiency of plasmid...frequency of base edits of interest than solely transfected cells. “Gene On” (GO)- a functional reporter ...

  7. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...interference (RNAi). The most common method is transfection of short interfering RNA (siRNA). Knockdown .... This approach often involves the transient transfection of expression plasmids in a given cell line....antibody should be able to detect even in cells transfected with low efficacy. But since there can be large...differences between endogenous expression and transfected overexpression, this approach cannot confirm ...

  8. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ... number of plasmids that are required in the transfection step of the viral packaging cell line. For example...resources on addgene.org Addgene Protocols: General Transfection The Addgene Retrovirus Guide  ...supplied by three types of plasmids, which are cotransfected into the 293T packaging cell line. This system...

  9. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...base pairs on each side, in fact. Later, when we transfect the cells, these large regions of homology enable...being cut and destroyed by CRISPR/Cas9 during transfection. Making the double strand break and introducing...

  10. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ... your packaging cell line, prepped your DNA, transfected and harvested your lentivirus. Now it’s time ...the correct transfer plasmid was used in the transfection; if preparing several different lentiviruses...

  11. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...ratio of acceptor to donor fusions varies with transfection efficiency and any unpaired fluorescent proteins...measure (overexpressing a gene or protein after transfection). Future of FRET The first genetically encoded...

  13. Viral Production at Addgene

    Type
    Blog Post
    ...the University of Pennsylvania Vector Core. Transfections are performed using the transfer plasmid, a ...primers targeting the transfer plasmid used in the transfection. PCR products are analyzed by gel electrophoresis...

  14. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...be produced through a single or dual-plasmid transfection, depending on whether the heavy and light chain...they’ve been separated onto two. You’ll first transfect your cells with your plasmid(s), and then wait...

  15. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...in mammalian cells is relatively quick. After transfection, HEK-293T cells start producing virus within...systems, it is standard to do only one passage (transfection). This differs from typical insect cell preparations...

  16. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...packaging and host genome integration. After co-transfection of these plasmids, 293T cells released transgene-containing...work with primary cells or other difficult-to-transfect cells. Lentiviral barcoding libraries have been...

  17. Zhang Lab CRISPR Page

    Type
    Collection
    ...human codon-optimized SpCas9; sgRNA can be co-transfected as PCR amplicons (Ran et al., Nature Protocols... #61422 - #61424 were designed for transient transfection : 61422 : Expresses dCAS9-VP64 activator with...codon-optimized SaCas9; SaCas9 sgRNA can be co-transfected as PCR amplicons using a protocol similar to ...

  18. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...identical targeting vectors delivered through transfection of naked DNA. Other investigators who had been...then packaged into AAV virions via transient transfection of 293T cells with helpers, and used to infect...

  20. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...site. (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences...efficacy of your gRNAs is then a simple matter of transfecting HEK293T cells with your modified pCAG-EGXXFP...
Showing: 41 - 60 of 145 results