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Genomic DNA
Reagent:
Genomic DNA
Default Quantity:
100 µg
Strains available:
H37Rv and Clinical isolate CSU93 (KY/TN 95-031551, CDC 1551)
Production system:
The bacilli are grown to late-log phase (day 14) in glycerol-alanine-salts (GAS) medium and harvested by centrifugation. Cell lipids are removed by chloroform/methanol (2:1) extraction in an equal volume of Tris-EDTA buffer. Delipidated cells are exposed to lysozyme and RNase overnight followed by SDS and Proteinase K treatment. Genomic DNA is isolated from contaminating proteins using organic extraction. To remove polysaccharide contaminants, DNA is precipitated with isopropanol and stored at 4°C as a dry pellet.
Notes:
Quality control for this reagent includes agarose gel electrophoresis and spectrophotometric analysis
References:
Belisle, J.T., and Sonnenberg, M.G., Methods Mol. Biol. 101:31, 1998.