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(Steve Chappell Mitchell, California Institute of Technology)
Mixture (each sample) 2.5µl 10X reaction buffer (Perkin Elmer) 2.5µl MgCl2 (25mM) 2.0µl dNTPs (2.5mM) 1.0µl vector primer 1 (5µM) 1.0µl vector primer 2 (5µM) 0.5µl AmpliTaq (Perkin Elmer) 14.5µl ddH2O
Cycles Time No. 94° 3min 1 94°(30sec)55°(30sec)72°(50sec) 30 72° 5min 1 4° hold
Reagent Final NaCl 1M Tris pH8.0 0.05M EDTA 5mM SDS 1% Dextran Sulfate 10%
Low SSC 1X SDS 0.1%
High SSC 0.1X SDS 0.1%