I use Chelex100 for extraction and MEGAquick-spinTotal for PCR purification. Some samples were very faint so i need to make salting out. Thanks in advance.
A half ml of blood sample was withdrawn from Jugular vein on EDTA tube as anticoagulant (0.2 ml of 0.5 M EDTA). DNA was freshly extracted from whole collected EDTA-blood. Two and half ml of lysis buffer TSTM (20 mM Tris-HCl pH 7.6, 640 mM sucrose, 2% Triton X-100, 10 mM MgCl2) was added to the aliquot. The mixture was centrifuged and the pellet suspended in 150 μl Proteinase K, 1.5 ml nuclei lysis buffer and 110 μl SDS 20%. After overnight incubation at 37◦ C, the proteins were removed by NaCl 6M and the DNA were precipitated by ice cold absolute ethanol. Extracted DNA was diluted with sterilized water in ratio 5:495μl before the optical density (OD),that is, purity and concentration of each DNA was carried out with the use of ultra violet spectrophotometer.