Kidd Lab 

SmaI site (RET18)

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

Primer 1.     RET18INTF:     5'-CCT GCA CAG CAG GAA AGT TC-3'                       100 ng/reaction
Primer 2.     RET18INTR:     5'-ACG ATG CAC ATC ATC ACG C-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

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"), 62^oC (30"), 72^oC (1')  × 30 cycles

Product size:
                                    Sma I
                   346 bp  -------------> 346 bp and 305 bp + 41 bp in heterozygote
 
 

Kidd Lab Histograms for Allele Frequencies