Saturday, May 15, 2004

Description
RBP3 BglII Polymorphism

Procedure
Bgl II polymorphism located near 5' end of intron 2

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

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 uM each

Buffer: Final Concentration 1.5 mM MgCl2, 50 mM KCl, 10 mM Tris HCl pH 8.4

Taq DNA Polymerase: 0.5 U/reaction
(Perkin Elmer Amplitaq DNA polymerase 5 U/ul, Cat. No. N808-0145)

PCR Cycling Profile: 95oC, 5'
95oC (30"), 61oC (30"), 72oC (1') x 30

Product Size:
Bgl II
400-----------> 310 + 90 (in homozygote site present)

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

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.


Recipes


Supplies


Tips