Saturday, May 15, 2004

Description
RET RsaI Polymorphism

Procedure
Rsa I Polymorphism in exon 15

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

Primer 1. RET15RSA1 5'-GAC TCG TGC TAT TTT TCC TCA3' 100 ng/Reaction
Primer 2. RET15RSA2 5'-TAT CTT TCC TAG GCT TCC CA 100 ng/Reaction

dNTPs: Final Concentration 200 M each

Buffer: Final Concentration 1.15 mM MgCl2, 50 mM KCl, 10 mM Tris HCl pH 8.4

Taq DNA Polymerase: 0.5 Units/reaction volume of 25 l
(Perkin Elmer Amplitaq DNA polymerase 5 U/l, Cat. No. N808-0145)

PCR Cycling Profile: 95oC, 5'
95oC (30"),562oC (30"), 72oC (30") 30 cycles

Product size: RsaI
260 bp----------> 140 bp and 120 bp (in homozygote site present)


Protocol from Ceccherini et al., 1994. Oncogene 9:3025-3029


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