Kidd Lab 

BglII site (located near 5' end of intron 2)

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

Primer 1.    RBP3BGL1:    5'-ACC CTG TGA CAC AGC AGA GG-3'        100 ng/reaction
Primer 2.    RBP3BGL2:    5'-ACC AGG GTT GTC ATG TGT TG-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"), 61^oC (30"), 72^oC (1')  × 30 cycles

Product Size:
                                  BglII
                   400 bp  ------------> 310 bp + 90 bp (in homozygote site present)

Note: On agarose gels stained with ethidium bromide, the 90 bp band is often not seen.

Reference:
Rodriguez, J.A., G.I. Liou, and S.P. Daiger, 1990. PCR detection of the Bglll RFLP at the RBP3 locus. Nucleic Acids Research 18:5578.
 
 

Kidd Lab Histograms for Allele Frequencies