Kidd Lab 

HaeIII site (located in exon 2)

Template: Genomic DNA 100-200 ng/reaction volume of 20 µl

Primer 1.     RET2HAE1:     5'-AGT GGC ATT GGG CCT CTA-3'               100 ng/reaction
Primer 2.     RET3HAE3:     5'-GCA GCC GTG TGC GGT ACG TG-3'        100 ng/reaction

dNTPs: Final Concentration 200 µM each

Buffer: Final Concentration 2.5 mM MgCl₂, 50 mM KCl, 10 mM Tris-HCl pH 8.4, 1 M Betaine

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"), 58^oC (30"), 72^oC (30")  × 30 cycles

Product size:
                                       HaeIII
                         170 bp ---------------->  106 bp +  63 bp (in homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies