M. leprae genomic DNA from three different isolates (NHDP-63, Br 4923, and Thai-53)
2 or 5 micrograms
Cell lipids are extracted from whole M. leprae bacilli
by chloroform/methanol (2:1) extraction in an equal volume of
Tris-EDTA buffer. Delipidated cells are exposed to 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 -70oC
as a dry pellet.
Quality control for this reagent includes agarose
gel electrophoresis, restriction enzyme digestibility, and spectrophotometric
Belisle, J.T., and M.G. Sonnenberg, Methods Mol.
Biol. 101:31, 1998.