Preparation of Template DNA by Boiling Method
| S. No. | Procedure |
|---|---|
| 1 | A single bacterial colony from overnight culture is suspended in 100 µL of 50 mM concentration of NaOH taken in eppendorf tube. |
| 2 | The tube is kept in a water bath at 97ºC for 3 minutes after properly sealing with paraffin tape. |
| 3 | Then the tube is placed into the refrigerator at 4ºC for 5 minutes. |
| 4 | After refrigeration, 16 µL of 1M of Tris-HCl is added and centrifuged at 8000 rpm for 2 minutes. |
| 5 | The supernatant is transferred and stored at -20oC. |
References
| S. No. | Papers |
|---|---|
| 1 | Hartas J, Hibble M, Sriprakash KS. Simplification of a locus-specific DNA typing method (Vir Typing) for Streptococcus pyogenes. J Clin Microbiol. 1998;36:1428-29. |
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