互联网2013-11-13
1. Vigorously mix the bottle of the HTR reagent for 30 seconds to ensure the particles are throughly resuspended. Add 100ìl of HRT reagent to 200 μl of sample and mix throughly by vortexing for 10 seconds.
Important: HTR reagent must be throughly suspended before being dispense from bottle. Tip: Use 1ml pipettor and cut off the end of 1ml tip to make it easier for pipetting the HTR reagent.
2. Incubate at room temperature for 2 minutes.
3. Centrifuge at full speed (>13,000 x g) for 2 minutes.
4. Transfer cleared supernatant to a new 1.5 ml tube. Note: If the supernatant still shows dark color from soil at this point, perform the HTR extraction again by repeating step 1-4.
5. Add 1 volume of isopropanol and mix by inverting 5-10 times
6. Centrifuge at 13,000 x g for 10 minutes to pellet DNA. Carefully decant the supernatant without disturbing the pellet
7. Wash the pellet in 70% ethanol.
8. Centrifuge at 13,000 g for 10 minutes. Carefully decant the supernatant without disturbing the pellet.
9. Let the pellet air dry for 5-10 minutes and resuspend in water or TE Buffer.
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