Saturday, May 15, 2004

Description
PAH AluI Polymorphism


Procedure
AluI; LOCATED IN EXON 7

Template: Genomic DNA 100-200 ng/reaction volume of 25 l

Primer 1. PAHALU333 5' CTTGCACTGGTTTCCGCCTC 3' 100 ng/reaction
Primer 2. PAHALU237 5' CCCAAACCTCATTCTTGCAGCA 3' 100 ng/reaction

dNTPs: Final Concentration 200 M each

Buffer: Final Concentration 1.50 mM MgC12, 50 mM KCl, 10 mM Tris HCl pH 8.4

Taq DNA Polymerase: 0.5 Units/reaction
(Perkin-Elmer Amplitaq DNA polymerase 5 U/l, Cat. No. N808-0145)

PCR Cycling Profile: 95C, 5'
95C (30"), 59C (30"), 72C (30") x 30 cycles

Product size: AluI
213 bp -----------> 124, 59 and 30 bp in AluI site present homozygote;
213 bp------------> 154 and 59 bp in AluI site-absent homozygote
NOTE: the 59 bp fragment is a constant fragment


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