Revision as of 12:41, 20 June 2007 by Luw134 (New page: Miniprep of Bacterial Genomic DNA #Grow bacterial strain to saturation #Spin 1.5 mL for 2 min in microcentrifuge #Resuspend in 567 ul TE buffere, 30ul of 20 mg/ml proteinase K. Mix and in...)
Miniprep of Bacterial Genomic DNA
- Grow bacterial strain to saturation
- Spin 1.5 mL for 2 min in microcentrifuge
- Resuspend in 567 ul TE buffere, 30ul of 20 mg/ml proteinase K. Mix and incubate 1 hr at 37C
- Add 100ul of 5M NaCl. Mix thoroughly
- Add 80 ul of CTAB/NaCl solution. Mix. Incubate 10 min at 65C.
- Extract with an equal volume of chloroform/isoamyl alcohol. Spin 5 min in microcentrifuge.
- Transfer aqueous phase to a fresh tube. Precipitate DNA with .6 vol isopropanol. Wash precipitate with 70% ethanol. Remove supernatant and briefly dry pellet in lyophilizer.
- Resuspend pellet in 100ul TE buffer.