Autoclave 5M sodium chloride and 13% (w/v) PEG 8000 prior to use.
1) Adjust the DNA solution to a final sodium chloride concentration of 0.8M and mix.
2) Add an equal volume of 13% PEG 8000 and mix. The volume of PEG should be the same as the volume of DNA + the volume of sodium chloride from step (1).
3) Place the sample in ice for 20 min.
4) Centrifuge at full speed for 15 min. at 4 C if you can or room temperature if you can’t.
5) Decant the supernatant and invert the tube on a clean paper towel or kimwipe with the lid folded back. Touch the rim of the tube to a dry portion of towel to remove as much supernatant as possible.
6) Add 0.5 mL of nuclease-free 70% ethanol to the tube, close the lid, and vortex for 5 seconds.
7) Centrifuge at full speed for 5 min. at 4 C or room temperature.
8 ) Decant the supernatant and invert the tube on a clean paper towel or kimwipe with the lid folded back. Touch the rim of the tube to a dry portion of towel to remove as much supernatant as possible.
9) Dry the pellet in a speedvac.
10) Resuspend the pellet in water or 10 mM Tris, 0.1 mM EDTA. Higher concentrations of EDTA will reduce the Mg concentration in the sequencing reaction and lead to crummy reactions. Adjust the DNA concentration to 250 ng/ mL before submitting samples for sequencing.