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We narrowed to 9 results for: expression vector

Showing: 1 - 9 of 9 results

  1. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...stable cell lines expressing a gene of interest from an integrated lentiviral vector....stable cell lines expressing a gene of interest from an integrated lentiviral vector. Unlike the short-term...using lentiviral vectors enables long-term protein expression studies. Moreover, repeating experiments ... the most desirable level of expression. Over time, transgene expression in a polyclonal population may...Lines with Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...short-term protein expression observed using transient transfection approaches, generating cell lines using...repeated transient transfection. Some lentiviral vectors deliver mammalian antibiotic resistance (e.g., ...

  2. 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...same orientation on your target vector. If you are not sure what vector to use, you can check out our Empty...Congratulations, you now have YGOI in a mammalian expression vector and can begin your studies. Reference Page...be used to easily move any piece of DNA from one vector to another as long as it is already bounded by ...of interest (YGOI for short). You might need to express YGOI in cultured mammalian cells. The problem is...orientation in the recipient plasmid. (You don't want to express the antisense version of your gene!) Are in frame...manufacturer’s instructions. Isolate your insert and vector by gel purification Run your digested DNA on an...

  3. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...TRC-Cloning Vector Click here to see the sequence of pLKO.1 TRC-cloning vector. The vector is 8901 base...-Cloning Vector I.2 Recipes I.3 Warranty information Back to Top A. pLKO.1-TRC Cloning Vector A.1 The ...use of this vector. A.2 Map of pLKO.1 pLKO.1 is a replication-incompetent lentiviral vector chosen by the...TRC-cloning vector. Addgene Plasmid Description pLKO.1 – TRC control Negative control vector containing... pLKO derived vectors that may also be useful for your experiment. To see these vectors, visit Addgene...and ligated into the pLKO.1 vector, producing a final plasmid that expresses the shRNA of interest. C.1...TRC Cloning Vector The following steps were the prevailing guidelines when the pLKO.1 vector was created...

  4. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...could lead to reduced transgene expression over time, as the lower expressing clones take over the polyclonal...by Limiting Dilution You may also like... Viral Vector Guides Virus Blog Posts Addgene Protocols Viral...that are more likely to retain stable transgene expression. Other methods of generating a monoclonal cell...population be monitored regularly for transgene expression. This protocol has been optimized for adherent...sufficiently expanded, screen the lines for transgene expression and/or other phenotypes. For example, perform...for lines with the highest or lowest transgene expression ( Figure 2 ). Sample Data Figure 1: Generation...from expansion of individual A549 cells stably expressing Cas9. A549 cells were transduced with lentiCas9...

  5. General Transfection

    Type
    Protocol
    ...General Transfection You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...polyethylenimine. Transfections allow for transient expression of a gene of interest in a target cell line and...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...

  6. Fluorescence Titering Assay

    Type
    Protocol
    ...Assay for Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...marker. Determining the titer of your lentiviral vector allows you to control the multiplicity of infection...protocol was developed using low titer lentiviral vectors (10 5 TU/mL). If you anticipate that your viral... transduction, cells were assayed for mCherry expression using fluorescence microscopy. Last reviewed ...

  7. Lentivirus Production

    Type
    Protocol
    ...Lentivirus Production You may also like... Viral Vector Guides Mol Bio Protocols Introduction This protocol...be used to produce lentivirus from a lentiviral vector transfected into 293T cells using a polyethylenimine...Figure 1: 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to μg PEI...

  8. Protocol - Bacterial Transformation

    Type
    Protocol
    ...plasmids (even those designed for mammalian cell expression) carry both a bacterial origin of replication...approximately 10-fold lower for ligation of inserts to vectors than for an intact control plasmid. Incubate the...

  9. AAV Production in HEK293 Cells

    Type
    Protocol
    ...Cells You may also like... AAV Viral Service Viral Vector Guides Virus Blog Posts Molecular Biology Reference...transfection: pHelper; pRC (Rep-Cap), plasmid expressing your gene of interest Triton X-100 Benzonase/...
Showing: 1 - 9 of 9 results