Kidd Lab 

CfoI site (locted 10 kb from MspI site)

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

Primer 1.     D3MSPSNPF:     5'-AGT GCA TGA TTT TGC CGT TTA G-3'        100 ng/reaction
Primer 2.     D3CFOR:            5'-CTT GGC ATG AGT GGT TTA GAC-3'          100 ng/reaction

dNTPs: Final Concentration 200 µM each

Buffer: Final Concentration 1.15 mM MgCl₂, 50 mM KCl, 10 mM Tris-HCl pH 8.4

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

PCR Cycling Profile: 94^oC( 5')
                              94^oC (30"), 58^oC (30"), 72^oC (1')  × 30 cycles

Product size:
                                CfoI
                   321 bp -----------> 243 bp + 78 bp (in homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies