2.7. Method 4
In this method sonication step was not included, instead, the ’snap-freeze-thaw’ method is replaced with snap-chill (at -80C) for 10 minutes and thaw (with higher temperature) at 95C (instead of 70C) for 5 minutes and this snap-chill-boil cycle repeated two more times with vortexing for 30 seconds between each cycle (Table 1). 10 µl of DNase-Free-RNase were spiked in the sample after cooling in ice for 2 minutes and incubated at 37ºC for 10 minutes to improve the quality of DNA, The remaining protocol was performed as per method 1.
(Table 1 here)