Hop DNA Isolation

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(Hop DNA Extraction Protocol)
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#Obtain an adequate amount of fresh hop leaves and crush them with liquid Nitrogen and a small amount of Carborundum powder (fine 320 grit).
#Obtain an adequate amount of fresh hop leaves and crush them with liquid Nitrogen and a small amount of Carborundum powder (fine 320 grit).
#Assume 90% of mass is water weight.
#Assume 90% of mass is water weight.
-
#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
+
#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.
 +
#Put 900μl into 6 tubes and add 600μl of 24:1 CHCl3:octanol to each and shake.
 +
#Centrifuge at 5000g for 10 minutes.
 +
 

Revision as of 14:37, 26 March 2007

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.




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.

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