Saturday, May 15, 2004

Description
RBP3 MspI Polymorphism

Procedure
MspI Polymorphism located near 3' end of intron 1

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

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

dNTPs: Final Concentration 200 uM

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"), 58oC (30"), 72oC (1') x 30

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


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


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