Kidd Lab 

MscI site

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

Primer 1.     D3BALIR:     5'-AAG TCT ACT CAC CTC CAG GTA-3'         100 ng/reaction
Primer 2.     D3BALIF:     5'-GCT CTA TCT CCA ACT CTC ACA-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 volume of 25 µl
                                 (Perkin-Elmer Amplitaq DNA polymerase 5 U/µl, Cat. No. N808-0145)

PCR Cycling Profile: 95^oC( 5')
                              95^oC (30"), 58^oC (30"), 72^oC (1')  × 30 cycles

Product size:
                               MscI
                   420 bp-----------> 304 bp + 111 bp + 47 bp (in homozygote site absent)
 
 

Kidd Lab Histograms for Allele Frequencies