Hop DNA Isolation

Hop DNA Extraction Protocol

1. Obtain an adequate amount of fresh hop leaves and crush them with liquid Nitrogen and a small amount of Carborundum powder (fine 320 grit).
2. Assume 90% of mass is water weight.
3. Add 3.3ml of buffer per gram of wet (16ml per gram of dried) hop leaves, and incubate for 1 to 4 hours at 60-65C.
4. Put 900μl into 6 tubes and add 600μl of 24:1 CHCl3:octanol to each and shake.
5. Centrifuge at 5000g for 10 minutes.
6. Remove supernatant and put in new vials (800μl in each).
7. Add 5μl of RNAase in each vial.
8. Add 5ml of buffer wash solution to each vial.

Buffer: 100ml: tris/HCl (ph=8.0) 50mM, NaCl 1.8M, EDTA 50mM. Then add 200mg/ml of CTAB and 20 μl/ml 2-mercaptoethanol. Buffer wash solution: