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VSVG Psudotyped Retrovirus Production
Published: Dec 5, 2011 DOI: 10.21769/BioProtoc.158 Views: 18102
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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
- Dulbecco's modified eagle medium (DMEM) (Life Technologies, Gibco®, catalog number: 11995-065 )
- Fetal bovine serum (FBS) (Gemini Bio-Products, catalog number: 900-108 )
- Penicillin/streptomycin solution (Life Technologies, Gibco®, catalog number: 15140-122 )
- NaCl (Sigma-Aldrich, catalog number: S7653 )
- HEPES (Sigma-Aldrich, catalog number: H7523 )
- Na2HPO4 (Sigma-Aldrich, catalog number: S7907 )
- CaCl2 (Sigma-Aldrich, catalog number: C5080 )
- Chloroquine (Sigma-Aldrich, catalog number: C6628 )
- Sodium butyrate (Sigma-Aldrich, catalog number: B5887 )
- Gag/pol (Cell Biolabs, catalog number: RV-111 )
- VSVG and retrovirus vector (Clontech, catalog number: 631512 )
- Calcium-phosphate (Ca-P) transfection solution (see Recipes)
- 2x HeBS (pH 7.0) (see Recipes)
- DMEM culture medium (see Recipes)
Equipment
- 24-Well plate
- Centrifuges
- Water bath
- 10 cm tissue culture plate
- 0.45 micro filter
Procedure
Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.
Category
Molecular Biology > RNA > Transfection
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