Kidd Lab 

RsaI site (located in exon 15)

Template: Genomic DNA 50-100 ng/reaction volume of 25 µl

Primer 1.     RET15RSA1:     5'-GAC TCG TGC TAT TTT TCC TCA-3'        100 ng/reaction
Primer 2.     RET15RSA2:     5'-TAT CTT TCC TAG GCT TCC CA-3'          100 ng/reaction

dNTPs: Final Concentration 200 µM 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
                                 (Perkin-Elmer Amplitaq DNA polymerase 5 U/µl, Cat. No. N808-0145)

PCR Cycling Profile: 95^oC (5')
                              95^oC (30"), 56^oC (30"), 72^oC (30")  × 30 cycles

Product size:
                                 RsaI
                   237 bp -------------->  108 bp +  129 bp (in homozygote site present)

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

Kidd Lab Histograms for Allele Frequencies