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VSVG Psudotyped Retrovirus Production

Jia Li Jia Li

Published: Dec 5, 2011 DOI: 10.21769/BioProtoc.158 Views: 18061

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Abstract

Retrovirus vector pseudotyped with vesicular stomatitis virus G (VSV-G) protein has been proven to exhibit high efficiency to deliver genes in a variety of cells. Efficiency is affected by relative cell growth rate and phosphatidylserine level on the cell membrane.

Materials and Reagents

  1. Dulbecco's modified eagle medium (DMEM) (Life Technologies, Gibco®, catalog number: 11995-065 )
  2. Fetal bovine serum (FBS) (Gemini Bio-Products, catalog number: 900-108 )
  3. Penicillin/streptomycin solution (Life Technologies, Gibco®, catalog number: 15140-122 )
  4. NaCl (Sigma-Aldrich, catalog number: S7653 )
  5. HEPES (Sigma-Aldrich, catalog number: H7523 )
  6. Na2HPO4 (Sigma-Aldrich, catalog number: S7907 )
  7. CaCl2 (Sigma-Aldrich, catalog number: C5080 )
  8. Chloroquine (Sigma-Aldrich, catalog number: C6628 )
  9. Sodium butyrate (Sigma-Aldrich, catalog number: B5887 )
  10. Gag/pol (Cell Biolabs, catalog number: RV-111 )
  11. VSVG and retrovirus vector (Clontech, catalog number: 631512 )
  12. Calcium-phosphate (Ca-P) transfection solution (see Recipes)
  13. 2x HeBS (pH 7.0) (see Recipes)
  14. DMEM culture medium (see Recipes)

Equipment

  1. 24-Well plate
  2. Centrifuges
  3. Water bath
  4. 10 cm tissue culture plate
  5. 0.45 micro filter

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Molecular Biology > RNA > Transfection

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