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We narrowed to 5 results for: Mammalian expression plasmids

Showing: 1 - 5 of 5 results

  1. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...bacterial expression vector. Using subcloning, you can easily move YGOI into a mammalian expression vector...YGOI for short). You might need to express YGOI in cultured mammalian cells. The problem is that the only...recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid Will...correct orientation in the recipient plasmid. (You don't want to express the antisense version of your gene...sites to your recipient plasmid : You can modify the MCS of your recipient plasmid using Annealed-oligo ...material. We recommend 1.5-2μg of donor plasmid and 1μg of recipient plasmid. It is also critical that as much...re-circularization of the recipient plasmid. You should treat your digested recipient plasmid with a phosphatase prior...

  2. General Transfection

    Type
    Protocol
    ...Use this protocol to transfect mammalian cells with your plasmid of interest....transfecting mammalian cells using linear polyethylenimine. Transfections allow for transient expression of a ...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ..., the plasmid should also be propagated in an endonuclease negative E. coli strain such as NEB stable....is prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the cells 1–2 ...the cells 24–48 h before checking for protein expression. Sample Data Legend: Lenti-X 293T cells were ...here by the amount of green fluorescent protein expression (green in the right panels) with a limited effect...

  3. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging plasmid 250 ng pMD2.G envelope plasmid to 20 μl serum-...to vary the ratio of shRNA plasmid, packaging plasmid, and envelope plasmid to obtain the ratio that gives...drives expression of puromycin. Puro R Puromycin resistance gene for selection of pLKO.1 plasmid in mammalian...shRNA. psPAX2 Packaging plasmid for producing viral particles. pMD2.G Envelope plasmid for producing viral... also be used with packaging plasmid pCMV-dR8.2 dvpr and envelope plasmid pCMV-VSVG from Robert Weinberg...into the pLKO.1 vector, producing a final plasmid that expresses the shRNA of interest. C.1 Recommended ... transient knockdown of protein expression, you may transfect plasmid DNA directly into the target cells...

  4. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ... the most desirable level of expression. Over time, transgene expression in a polyclonal population may...Protocol to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector...protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral...lentiviral vector. Unlike the short-term protein expression observed using transient transfection approaches... lentiviral vectors enables long-term protein expression studies. Moreover, repeating experiments in a...transfection. Some lentiviral vectors deliver mammalian antibiotic resistance (e.g., puromycin, blasticidin...check the cells regularly to confirm transgene expression. As polyclonal populations of resistant cells...

  5. Protocol - Bacterial Transformation

    Type
    Protocol
    ...replicating plasmids. Because of this, nearly all plasmids (even those designed for mammalian cell expression... also like... Plasmid Cloning by Restriction Enzyme Digest Restriction Digest of Plasmid DNA DNA Ligation...that will help to maintain the plasmid without rearrangement of the plasmid DNA. Additionally, specific ...tube of plasmid DNA and just need to transform bacteria so that you can grow up more of the plasmid) you ...efficient at taking up larger plasmids. If you need to transform large plasmids, it is a good idea to use ...Learn how to transform E. coli with your plasmid of interest. Protocols...introduced into a cell. Transformation of bacteria with plasmids is important not only for studies in bacteria ...
Showing: 1 - 5 of 5 results