Hey everyone,

I have a bacteria project for my masters, but isolating the chemistry from them (my background is chemistry). I have cell cultures in TSB, took an aliquot in an eppendorf, centrifuged to concentrate the cells at the bottom and removed the supernatant. I have an SOP for extracting the DNA, but I am not sure if it is working out correctly, could the experts here let me know? I'll consolidate it:

1. add 1X TE Buffer
2. remove supernatant and add 1.6% sarkosyl and proteinase K (to supernatant)
3. incubate at 60C for 1 hour
4. add PCI and centrifuge
5. remove aq layer and add 100% IPA and 3M sodium acetate (to supernatant)
6. freeze in -80C for 30 min; centrifuge
7. discard supernatant
8. add 70% EtOH to pelleted DNA and centrifuge; discard supernatant
9. air dry sample, then resuspend in molecular grade water
10. measure concentration on NanoDrop

Any help is greatly appreciated!!