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*Pellet at full speed in a microfuge, and aspirate supernatant.
*Add 1.8ml Lysis Buffer to 0.2ml 10X Lysozyme/RNAse for every ten samples.
10X Lysozyme/RNAse:
(Store @ -20degC)
5mg/ml Lysozyme
1mg/ml RNAse (Boiled)
Lysis Buffer:
(Store @ 4degC)
25ml EDTA (500mM)
62.5 Triton X-100 (20%)
62.5 Sucrose (32%)
12.5 Tris-HCl pH8 (1M)
87.5 Water
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250
*Add (150) 200ul Lysis Buffer +Lysozyme/RNse to pelleted cells and vortex to mix.
*Leave at RT for 10min.
*Boil for 2min.
*Cool on ice.
*Spin @ full speed in a microfuge for 30min.
*Toothpick out pelleted debris.
*Add (150) 200ul isopropanol to remaining supernatant and vortex to mix.
*Spin @ full speed for 10min in a microfuge.
*Aspirate isopropanol and resuspend pellet in 50ul TE or water.