Kidd Lab 

MspI site ( located in RET14)

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

Primer 1.     RETRESQ14F :     5'-GGA AGC GCA GGG CAC TAC G-3'           100 ng/reaction
Primer 2.     RETRESQ14R:     5'-AGG TGC AGC AGG TCA AAT ACG-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"), 61^oC (30"), 72^oC (1.5')  × 30 cycles

Product size:
                                   MspI
                   1092 bp ------------> 800 bp and 600 bp + 200 bp in heterozygote (with a constant band of 290 bp)
 
 

Kidd Lab Histograms for Allele Frequencies