Kidd Lab 

MspI site

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

Primer 1.     S94MSPA:     5'-CGA ATT CTT CCT GAA GCC T-3'          90 ng/reaction
Primer 2.     S94MSPB:     5'-TTT GTA ATG GAG TGG TAT CT-3'        90 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
                                 (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 (30")  × 30 cycles
                              72^oC (7')

Product size:
                                  MspI
                    307 bp--------------> 190 bp + 117 bp (in homozygote site present)

Reference:
Brooks-Wilson, A.R., D. Smailus, D. Gilchrist, and P.J. Goodfellow, 1992.  Additional RFLPs at D10S94 and the development of PCR-based variant detection systems: Implications for disease genotype prediction in MEN2A, MEN2B, and MTC1 families.  Genomics 13: 233-234.
 
 

Kidd Lab Histograms for Allele Frequencies