DNA Extraction - Qiagen DNeasy Kit
DNA Extraction – Qiagen DNeasy kit.
Note the number of spin columns you use.
The centrifuge can hold up to 30 samples at once.
Forceps are to be sterilized in Fine Science Tools heat block at >200 C for>20 seconds.
1. Isolate a suitable piece of tissue and place in a UV-crosslinked 1.5mL tube.
2. Add 180uL Buffer ATL and 20uL Proteinase K and vortex.
3. Place in the 55 C incubator for 3 hours or overnight.
4. Remove from incubator, vortex, add 200uL Buffer AL and vortex.
5. Place in heat block at 70 C for 10 minutes.
6. Add 200uL 100% Ethanol and transfer entire volume onto spin column.
7. Centrifuge at 8000 rpm for 1 minute; discard flow-through.
8. Add 500uLBuffer AW1 and centrifuge at 8000 rpm for 1 minute; discard flow-through.
9. Add 500uL Buffer AW2 and centrifuge at 13000 rpm for 3 minutes; discard flow through.
10. Place spin column on UV-crosslinked 1.5mL tube, add 200uL buffer AE. Let sit for 1 minute, then centrifuge at 8000 rpm for 1 minute. Repeat and then combine flow-throughs for a total volume of 400uL.