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Method: Restriction Digests of High Molecular Weight Yeast DNA

Jim Howe

May 5, 1990


Purpose:

Time required:

Special Reagents:

Procedure:

  1. Using wide-bore pipette, aliquot 100 ng of liquid DNA into eppendorf tube. Add 2.5 Ál 10X restriction buffer, 1-2 ul RNase (5 mg/ml stock), 250-500 ng BSA, then dH2O up to 25 Ál volume. Incubate at appropriate temperature for 2-4 hours.
  2. Stop the reaction by adding 3 Ál of 10X glycerol stop mix; mix in tube gently, then microfuge for 15 seconds (rather than pipette up and down, which may shear the DNA).