Kidd Lab 

MspI site (located near 3' end of intron 1)

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

Primer 1.    RBP3MSP3:    5'-CTG TAA GAG TTG GAG GT-3'             100 ng/reaction
Primer 2.    RBP3MSP1:    5'-GTG CTT CAG TTT TTC CAT GG-3'        100 ng/reaction

dNTPs: Final Concentration 200 µM

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"), 55^oC (30"), 72^oC (1')  × 30 cycles

Product Size:
                                  MspI
                   515 bp  ------------> 394 bp + 121 bp (in homozygote site present)

Reference:
Wu, J. and M. Nickerson, 1991. PCR detection of the MspI polymorphism in the human IRBP gene (RPB3). Nucleic Acids Research 19:4016.
 
 

Kidd Lab Histograms for Allele Frequencies