Proteinase K method for DNA extraction protocol
PROTOCOL 1) In 2 ml Eppendorf Safe-Lock tube with precisely upset creature or plant tissues include new 500 μl of extraction cradle (0.8 M guanidine thiocyanate, 10 mM EDTA, 5% Tween 20, 0.5% Triton X-100, 50 mM HEPES-acid*) with 200 μg of proteinase K. 2) vortex quite well and hatch the examples at 55°C for a few hours or better for the time being at 37°-55°C (the more drawn out the better, until disintegrate tissue) with infrequent vortexing. 3) Include 700 μl of chloroform, vortex very well for 1 moment making an emulsion (in the MM300 Mixer Mill at 30 Hz); alternatively: brood the examples at 55°C during 30 min. 4) Turn at most extreme speed in a microcentrifuge for 5 minutes. 5) Move the supernatant into another 2 ml tube containing 500 μl of 2-propanol and 100 μl 3M Na-acetic acid derivation, vortex well overall, and rotator the cylinders at most extreme speed in a microcentrifuge for 4 minutes. 6) Dispose of the supernatant and include 1.8 ml of 70% ethano...