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Rapid DNA Extraction from Cyanobacteria

Author: Sanchita Sinha, Pawan K. Dadheech and M. KrishnaMo
Date Added: Mon Feb 02 2009
Date Modified: Mon Feb 02 2009

Materials

Procedure

  1. Macerate Cyanobacteria in TE Buffer (10T/1E)
  2. Pellet the cells by centrifugation for 2 min at 10,000 rpm at room temperature
  3. Remove supernatant. Add 500 l TE buffer (10T/1E).
  4. Add 1% SDS and 50 l of 50 mg/ml lysozyme stock solution
  5. Keep at 70C for 15 min.
  6. Add equal volume of Phenol : Chloroform
  7. Centrifuge for 10 min at 10,000 rpm at room temperature
  8. Take supernatant and extract it twice using 100% Chloroform extraction procedure.
  9. Finally take supernatant and add 0.1 volume of 5M NaCl and 2 volumes of 100% ethanol and keep for precipitation at 20C for 2 hrs
  10. Centrifuge the tubes for 30 min at 15,000 rpm at 10C
  11. Wash the pellet twice by 70% ethanol by centrifuging it at 10,000 rpm for 10 minutes.
  12. Dry the pellet and resuspend in TE buffer (10T/1E)
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