Biochemical Techniques and Instrumentation

Materials

1. Genomic DNA extraction buffer (250ml):
   1M Tris HCl (pH 8.0) - 2.5ml
   0.5M EDTA (pH 8.0) - 50 ml
   Pancreatic RNase - 5 mg
   10% SDS - 12.5 ml
   Adjust pH to 8.0 and adjust volume to 250ml with double distilled water
2. Saturated phenol (pH 8.0) : Mix equal amounts of 0.5M Tris HCl (pH 8.0) buffer and phenol. Allow it to separate in a separating funnel overnight in a dark place. Collect the lower layer containing the saturated phenol. Store in a amber colored bottle.
3. 10M ammonium acetate (NH₄Ac)
4. Proteinase K (20mg/ml stock)
5. Ethanol, 100%
6. TE buffer (10X):
   1 M Tris-HCl (pH 7.5) - 100 ml
   0.5M EDTA (pH 8.0) - 20 ml
   Double distilled water - 880 ml