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Purification of Adenovirus by Cesium Chloride Density Gradients
Published: Apr 5, 2012 DOI: 10.21769/BioProtoc.201 Views: 34282
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Abstract
Adenovirus are efficient gene delivery systems. The standard method for purification of adenoviral vectors is based on using a cesium chloride (CsCl) density gradient combined with ultracentrifugation. This method is suitable for small-scale purification and is less expensive than column chromatography or commercial purification kits.
Background
Materials and Reagents
- HEK293 cell
- Glycerol
- Liquid nitrogen
- 70% ethanol
- Distilled water
- SDS
- CaCl2
- MgCl2
- CsCl2 (Thermo Fisher Scientific, catalog number: BP1595-500 )
- Tris-HCl (pH 8.0)
- TE
- EDTA
- SDS
- Sucrose
- 5% deoxycholate (see Recipes)
- Dialysis buffer (see Recipes)
- Balance buffer (see Recipes)
- Saturated CsCl2 (see Recipes)
Equipment
- Centrifuges
- Ultracentrifuge
- Laminar flow hood
- Syringe
- Water bath
- Slide-A-Lyzer 10 K dialysis cassette (Thermo Fisher Scientific, catalog number: 66203 )
- Spectrometer
- Ti50.2 tube
Procedure
Category
Microbiology > Microbial cell biology > Cell isolation and culture
Microbiology > Microbe-host interactions > Virus
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