Saturday, May 15, 2004

Description
D10S94 PvuII Polymorphism

Procedure
Pvu II

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

Primer 1. S94PVUA 5 -AGCTCACCCAGATCCTAA -3' 90 ng/reaction
Primer 2. S94PVUB 5' -CCTCCCTTCAGTAGACAC -3' 90 ng/reaction

dNTPs: Final Concentration 200 M each

Buffer: Final Concentration 1.5 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"), 54C (30"), 72C (30") x 30 cycles

Product size: Pvu II
211 bp ----------->145 bp and 66 bp (in homozygote site present)

Ref: Brooks-Wilson, A.R., D. Smailus, D. Gilchrist, and P.J. Goodfellow, 1992. Additional RFLPs at D10S94 and the development of PCR-based variant detection systems: Implications for disease genotype prediction in MEN2A, MEN2B, and MTC1 families. Genomics 13: 233-234.


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