Kidd Lab 

BstNI site (located 10 kb from the MspI site)

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

Primer 1.     D3BSTNIF:     5'-CTA GGC TCT GAA AAG AAA TGT CA-3'        100 ng/reaction
Primer 2.     D3BSTNIR:     5'-TGC TTA ATT GTC TTC ACC TCA C-3'             100 ng/reaction

dNTPs: Final Concentration 200 µM each

Buffer: Final Concentration 1.15 mM MgCl₂, 50 mM KCl, 10 mM Tris-HCl pH 8.4

Taq DNA Polymerase: 0.5 Units/reaction volume of 25 µl
                                 (Perkin-Elmer Amplitaq DNA polymerase 5 U/µl, Cat. No. N808-0145)

PCR Cycling Profile: 94^oC( 5')
                              94^oC (30"), 56^oC (30"), 72^oC (1')  × 30 cycles

Product size:
                                BstNI
                   303 bp---------------> 222 bp + 81 bp (in homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies