Kidd Lab 

SacI 5' G-355A site (located in 5' promoter region)

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

Primer 1.     TDI_EX12.1:     5'-CGA GTA CGG GCT GCA GGC ATA CAC T -3'            100 ng/reaction
Primer 2.     EX12R4 :         5'-CCC AGC AGG TCC CAC AAT CAC AG-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: 94^oC (5')
                              94^oC (30"), 59^oC (30"), 72^oC (30")  × 30 cycles
                              72^oC (10')

Product size:
                                   SacI
                    133 bp--------------------> 111 bp + 22 bp (in homozygote for site present)
 
 

Kidd Lab Histograms for Allele Frequencies