Kidd Lab 

EcoRI site

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

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 µM/reaction

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

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

PCR Cycling Profile: 95^oC (5')
                              95^oC (30"), 62^oC (30"), 72^oC (1')  × 30 cycles

Product size:
                                 EcoRI
                   924 bp ---------------> 924 bp and 571 bp + 353 bp (in homozygote for site present)

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

Kidd Lab Histograms for Allele Frequencies