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  1. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ... 2019 Estimate of time required: ~3 h Workflow Timeline Plate set-up: 2 h qPCR run: 1.5 h Data analysis...°C 30 sec / read plate/ repeat 39x from step 3 / melt curve Example of plate set-up: 1 2 3 4 5 6 7 8 A...accurately determine differences between samples. Melt curve analysis: a single peak should be seen. The...

  2. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...Set annealing temperature 5°C below the primer melting temperature (Tm). Set extension step at 1-2 minutes... 3’ primers should be designed to have similar melting temperatures (Tm). Set the annealing temperature...

  3. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ... Qiaquick gel extraction kit Qiagen #28704 Low melting point agarose Sigma #A9414 Luria Broth Agar (LB...37°C for 2 hours. Run digested DNA on 0.8% low melting point agarose gel until you can distinctly see ...

  4. Protocol - How to Design Primers

    Type
    Protocol
    ...60% G/C content Start and end with 1-2 G/C pairs Melting temperature (Tm) of 50-60°C Primer pairs should...

  5. Ligation Independent Cloning

    Type
    Protocol
    ...Use web-based primer design software to ensure a melting temperature between 50-60°C for your PCR primers...

  6. Fluorescence Titering Assay

    Type
    Protocol
    ... institution’s biosafety regulations. Workflow Timeline Day 0: Seed 293T cells Day 1: Transduce cells ...

  7. General Transfection

    Type
    Protocol
    ... and different transfection reagents. Workflow Timeline Day 0: Seed HEK293T cells (or a subclone of HEK293T...

  8. Immunocytochemistry

    Type
    Protocol
    ...science. Last Update: January 20, 2022 Workflow Timeline Day 1: Seed cells Day 3-4: Fix and label cells...

  9. Lentivirus Production

    Type
    Protocol
    ... such as stable-cell line generation. Workflow Timeline Day 0: Seed 293T packaging cells Day 1 (pm): Transfect...

  10. Colony Formation Titering Assay

    Type
    Protocol
    ...cannot be used for later experiments. Workflow Timeline Day 0: Seed and transduce cells Day 2: Replace...

  17. Western Blot

    Type
    Protocol
    ...approaches. Last Update: January 24, 2022 Workflow Timeline Day 1: Prepare lysates, run SDS-PAGE, transfer...

  20. AAV Production in HEK293 Cells

    Type
    Protocol
    ...handle a large number of T-175 flasks. Workflow Timeline Day 0: Seed cells in CS2 Day 2: Seed cells in ...
Showing: 1 - 20 of 22 results