Saturday, May 15, 2004

Description
APOB EcoRI Polymorphism

Procedure
EcoR I

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

Primer 1. APOBEC1 5'-CTG GCT TGC TAA CCT CTC TG-3' 100 ng/Reaction
Primer 2. APOBEC2 5'-GAG AAG CTT CCT GAA GCT CG-3' 100 ng/Reaction

dNTPs: Final Concentration 200 M each

Buffer: Final Concentration 2.25 mM MgCl2, 50 mM KCl, 10 mM Tris HCl pH 8.4

Taq DNA Polymerase: 0.5 Units/reaction volume of 25 l
(Perkin Elmer Amplitaq DNA polymerase 5 U/l, Cat. No. N808-0145)

PCR Cycling Profile: 95oC, 5'
95oC (30"), 62oC (30"), 72oC (30") 30 cycles

Product size:
EcoRI
1000bp ----------> 1000bp and 600 bp + 400 bp in heterozygote

Boerwinkle, E., S.S.Lee, R. Butler, VN Schumaker, and L. Chan, 1990. Rapid typing of apolipoprotein B DNA polymorphisms by DNA amplification. Athero 81:225-232.


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