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1) add 250 µl plasma to 750 µl Triazole
2) 5 min incubation
3) add 150 µl chloroform, mix 12 sec, 3 min incubation
4) Centrifuge at 10,000g for 10 min at room temperature
5) Transfer the upper clear phase to a fresh tube
6) Add 0.5 mL of isopropanol per each milliliter of the clear phase
7) Mix vigorously by rapid shaking and 10 min incubation
8) Centrifuge at 10,000g for 10 min at 4°C
9) Remove any remaining liquid
10) wash with 500 µl 75% ethanol
11) Add 20 µl DEPS water
12) Dry bath, 10 min, 60°C
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