Kidd Lab 

SacI site

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

Primer 1.     MCADSCU2:     5'-TTT ACC TTT ATT TCT ATT GTG ATG-3'            100 ng/reaction
Primer 2.     MCADSCU4:     5'-TCT TTA AAG AAA ACT GTG TGT ACC-3'         100 ng/reaction

dNTPs: Final Concentration 200 µM each

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

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

PCR Cycling Profile: 95^oC (5')
                              95^oC (30"), 55^oC (30"), 72^oC (30")  × 30 cycles

Product size:
                                   SacI
                    313 bp-------------> 200 bp + 113 bp (in homozygote site present)

Modified from Zhang, Z., S. Kolvraa, Y. Zhou. D.P. Kelly, N. Gregersen, and A.W. Strauss, 1933. Three RFLPs defining a haplotype associated with the common mutation in human Medium-Chain Acyl-CoA Dehydrogenase (MCAD) deficiency occur in Alu repeats. Am. J. Hum. Genet. 52: 1111-1121.
 
 

Kidd Lab Histograms for Allele Frequencies