Kidd Lab 

DdeI site (RET15)

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

Primer 1.     Retresq15f2:     5'-GAG CAC TGC TTG TCC TGT CC-3'                       100 ng/reaction
Primer 2.     Retresq15r2:     5'-CCA TCT CCC ATG ATA CTC TCC-3'                      100 ng/reaction

dNTPs: Final Concentration 200 µM each

Buffer: Final Concentration 1.5 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"), 61^oC (30"), 72^oC (1')  × 30 cycles

Product size:
                                    DdeI
                   300 bp  -------------> 300 bp and 250 bp + 50 bp in heterozygote
 
 

Kidd Lab Histograms for Allele Frequencies