Kidd Lab 

BsrI site (located 10 kb from the MscI site toward the MspI site)

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

Primer 1.     D3MSCSNPF:     5'-CAG AGG CCT AGA GAA ACA CC-3'                100 ng/reaction
Primer 2.     D3MSCR2:          5'-AAG GAA AGG AAG GAC AGA TGC T-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"), 58^oC (30"), 72^oC (1')  × 30 cycles

Product size:
                                 BsrI
                   414 bp--------------> 296 bp + 118 bp (in homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies