Kidd Lab 

HindIII site (PCR location in AC006487: 804-1295 bp; HindIII location in AC006487: 969-974 bp)

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

Primer 1.     NGFRHND3F:    5'-CCC CTT GCG GAT GTT CTC-3'           90 ng/reaction
Primer 2.     NGFRHND3R:    5'-GTG GGA TGG CTG GAC CTG-3'        90 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"), 58^oC (30"), 72^oC (30")  × 30 cycles
                               72^oC (7')

Product size:
                                 HindIII
                   492 bp ---------------> 324 bp + 168 bp (in heterozygote)
 
 

Kidd Lab Histograms for Allele Frequencies