Materials 21 Reagents

1. CHAPS lysis buffer: 10 mM Tris-HCl pH 7.5, 1 mM MgCl2, 1 mM EGTA, 10% glycerol, 0.1 mM 4-amidinophenylmethane sulfonylfluoride (APMSF), 5 mM 0-mercaptoethanol, CHAPS. From 1 M stock solutions of Tris-HCl, pH 7.5, and MgCl2, 100% glycerol and solid EGTA, prepare lysis buffer without 0-mercaptoethanol, CHAPS, and APMSF. Autoclave, cool, then add CHAPS, 0-mercaptoethanol, and APMSF (50 mM stock in water). All the chemicals were

1 st PCR Cycle

Amplifluor ® Primer 1

Primer 2

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