Rapid DNA Extraction from Cyanobacteria
关键词: rapid dna extraction来源: 互联网
- Phenol
- Chloroform
- Absolute ethanol
- 70% ethanol
- TE buffer (10T/1E), pH 8.0
- 10% SDS
- 50 mg/ml lysozyme
- 5 M NaCl
Procedure
Macerate Cyanobacteria in TE Buffer (10T/1E) Pellet the cells by centrifugation for 2 min at 10,000 rpm at room temperature Remove supernatant. Add 500 µl TE buffer (10T/1E). Add 1% SDS and 50 µl of 50 mg/ml lysozyme stock solution Keep at 70°C for 15 min. Add equal volume of Phenol : Chloroform Centrifuge for 10 min at 10,000 rpm at room temperature Take supernatant and extract it twice using 100% Chloroform extraction procedure. Finally take supernatant and add 0.1 volume of 5M NaCl and 2 volumes of 100% ethanol and keep for precipitation at �20°C for 2 hrs Centrifuge the tubes for 30 min at 15,000 rpm at 10°C Wash the pellet twice by 70% ethanol by centrifuging it at 10,000 rpm for 10 minutes. Dry the pellet and resuspend in TE buffer (10T/1E)
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