Biotechnology and Bioinformatics (1+1)

Procedure

1. Collect the blood in ethanol, store in -20°C.

2. Mix well, take 100μl and add 100 μl of TE (Tris EDTA solution, pH 8.0).

3. Centrifuge at 3000rpm for 5 min.

4. Remove the supernatant and add 200 μl of TKEM buffer (10mM Tris-HCl,

pH 7.6, 10mM KCl, 2mM EDTA containing 4mM MgCl₂ )

5. Centrifuge at 3000rpm for 5 min. Remove the supernatant and add 200 μl TKEM and again centrifuge and remove the supernatant.

6. Add 200 μl of TKEM buffer and add 15 μl of 10% SDS. Mix well, keep at 55°C for 5 min.

7. Add 75 μl of 6M NaCl. Mix well, centrifuge at 10,000 rpm for 5 min.

8. Collect the supernatant and add ethanol to precipitate the DNA. The DNA will appear as threads. Keep overnight at -20°C.