DM1 Alu Polymorphism
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.
(N.B. This protocol gives
rather erratic results. Be forewarned, we do not recommend it but have
no other.)
Kidd Lab Histograms for Allele Frequencies
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