Saturday, May 15, 2004

Description
DM1 Alu Polymorphism

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

Primer 1: 405: 5'-CTG TAT ACT CAG CTA CTA GGG T-3' 50 ng/rx
Primer 2: 491: 5'-AAA TAG GCT GGA CCG CGG-3' 50 ng/rx
Primer 3: 486: 5'-CTC AGG GGT TAT CTA AAG TGG C-3' 100 ng/rx

dNTPs: Final Concentration 200 uM

Buffer: Final Concentration 50 mM KCl 10 mM Tris-HCl (pH8.4)
1.5 mM MgCl2

Taq DNA Polymerase: 0.5 U/reaction
(Perkin Elmer Amplitaq DNA Cat. No. N808-0145)

PCR Cycling Profile: 95oC, 5'
95oC (1'), 60oC (1.5'), 72oC (1.5') x 30

Product size: 1008 bp and 494 bp for heterozygous (+/-) sample

Protocol from: Mahadevan MS, Foitzik MA, Surh LC, and Korneluk RG, 1993. Characterization and Polymerase chian reaction (PCR) detection of an Alu deletion polymorphism in total linkage disequilibrium with myotonic dystrophy. Genomics 15:446-448.


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