Kidd Lab 

SpeI site (located in intron 14)

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

Primer 1.     NCAMINT14F2:     5'-CTT GCT GGG GTT TCG TAG TAG-3'        100 ng/reaction
Primer 2.     NCAMINT14R:      5'-GAG AGG GTG GTT ACG GTT GT-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"), 55^oC (30"), 72^oC (1')  × 30 cycles
                               72^oC (10')

Product size:
                                    SpeI
                   839 bp  ----------------> 495 bp + 58 bp (in homozygote for site present)
 
 

Kidd Lab Histograms for Allele Frequencies