Kidd Lab 

MspI (B) site (located about 8 kb centromeric of HindIII (B) site)

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

Primer 1.     G8SNPF:     5'-CTAGAAGGTGTAAGTCAATGAG-3'        100 ng/reaction
Primer 2.     G8MSPB:    5'-CCCATGCATTTTCCTTTTGAGC-3'           100 ng/reaction

dNTPs: Final Concentration 200 µM/reaction

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"), 54^oC (30"), 72^oC (45")  × 30 cycles

Product size:
                                 MspI on 3% gel
                    164 bp-----------------------------> 130 bp + 34 bp (with 31 bp constant site for homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies