Rapid Isolation of Genomic DNA from Human Oral Mucosa

Procedure

1. Take an oral smear
2. Dissolve in 50 ml H₂O through vigorous shaking
3. Pellet cells 5 min at 4000 rpm
4. Re-suspended the pellet in 400 μl Lysis buffer (50 mM TRIS-Cl pH 8, 10 mM EDTA, 2% SDS)
5. Incubate for 5 min at 65¡ãC
6. Add 250 μl 4.5 M NaCl and mix
7. Pellet cellular debris (13000 rpm for 4 min). Genomic DNA stays in the supernatant
8. Precipitate DNA by adding 650 μl of isopropanol to the supernatant and centrifuge 5 min at 13000 rpm
9. Re-suspend the DNA in an appropriate buffer (e.g. TrisCl, pH=8.0)

Reference
M. Ralser, R. Querfurth, H.J. Warnatz, H. Lehrach, M.L. Yaspo and S. Krobitsch An efficient and economic enhancer mix for PCR
BBRC, 2006. doi:10.1016/j.bbrc.2006.06.151