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1. Add 100 ¦Ìl of Solution A to the tail clip (2mm)

  2. Place in heat block (preheated to 95¡ãC) for 60 minutes

  3. Add 100 ¦Ìl of Solution B

  4. Vortex briefly

  5. Centrifuge at 2000rpm for 10 minutes

  6. Remove 100 ¦Ìl of supernatant and store in new labeled tube at 4¡ãC *

  7. Use 1¦Ìl per 25 ¦Ìl PCR reaction



* Optional-1 ¦Ìl of supernatant can be removed directly from samples and added to pcr reaction.



Solution A: 25mM NaOH/0.2mM EDTA



Solution B: 40mM Tris-HCl



Solutions A & B should be made up fresh each week, can be stored 1 week at 4¡ãC



STOCK SOLUTIONS:

  1. 5M NaOH

  2. 0.5M EDTA, pH 8.0

  3. 1M Tris-HCl pH 8.0
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