Gram pozitif bakterilerden genomik DNA izolasyon protokolü

Solüsyon ve kullanılan malzemeler

1. Lysis buffer: 75 mM sodium chloride, 25 mM EDTA, 20 mM Tris–HCl, pH 7.5, containing 10 mg/mL of lysozyme (This solution should be prepared just prior to use).
2. 10% sodium dodecyl sulfate (SDS).
3. 25 mg/mL proteinase K in distilled water.
4. 5 M sodium chloride.
5. 24 : 1 (v/v) mixture of chloroform and isoamyl alcohol.
6. 100% isopropanol.
7. 70% ethanol chilled to –20°C.
8. TE buffer: Tris–HCl 10 mM, and EDTA 1 mM, pH 8.0. Autoclave for 20 min at 121°C.
9. TE-buffer-saturated phenol.
10. 3 M sodium acetate adjusted to pH 5.2 with glacial acetic acid.
11. 37°C water bath.
12. 55°C water bath.
13. Refrigerated centrifuge.
14. pH Meter.

Agarose Gel Electrophoresis for DNA sizing.

Protocol according to M. Andrea Azcárate-Peril and Raúl R. Raya.