Kidd Lab 

MspI (A) site (located 13.3 kb upstream (telomeric) of HindIII (A) site)

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

Primer 1.     H552MSPR:    5'-GCT CCC CTC GCT AAC CTC-3'          100 ng/reaction
Primer 2.     H552BK1:       5'-CTC TGT CAA GCA TCC TCT G-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"), 55^oC (30"), 72^oC (30")  × 30 cycles

Product size:
                                 MspI
                    498 bp--------------> 275 bp + 223 bp (for homozygote site present)

Reference: Skraastad et al., Am. J. Hum. Genet. 44: 560-566 (1989); converted to PCR in Kidd Lab by sequencing.
 
 

Kidd Lab Histograms for Allele Frequencies