DNA Extraction from Bacteria:
1. Add 1.5ml broth in an epi and centrifuge it at 1000rpm for 10 min.
2. Discard the supernatant and add 400µl lysis buffer. Vortex it for 5 min.
3. Add 200µl of 10% SDS and mix it by inverting several times.
4. Place it in water bath at 65°c for 10 min.
5. After that place it in -20°c for 10 min to lysis cells through the heat shock.
6. Add 500µl of phenol chloroform isoamyl (PCI) and invert it few times.
After that centrifuge it for 10 min at 1000rpm. Pick the upper layer and put it
in new epi.
7. Repeat the above step.
8. After transferring upper layer in new epi add isopropanol (ice cold) and
place it at -20°c overnight.
9. Next day centrifuge it for 10 min at 1000rpm. Pellet will be visible. Discard
the supernatant.
10.Add 70% ethanol for washing the pellet and centrifuge it for 10 min.
11.Discard the supernatant and dry the pellet at room temperature by inverting
epi on tissue paper.
12.After that add 25µl TE buffer (1X) for dilution purpose and store it at -20°c.
DNA Lysis Buffer:
 5ml tris (1mM)
 10ml EDTA (0.5mM)
 2.5ml SDS (10%)
 0.1 ml NaCl (5mM)
 32.4ml autoclaved water