Kidd Lab 

HincII site (PCR location in AC006487: 11430-12116 bp; HincII location in AC006487: 11499-11504bp; 1695 bp downstream from exon1 of NGFR)

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

Primer 1.     NGFRHNCF:    5'-CAC CTT TTC AGG GGG CAG C-3'        100 ng/reaction
Primer 2.     NGFRHNCR:    5'-CCC TCG TTC CAT CCC TAT TA-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, 1M betane
[NOTE THE BETANE]

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 (1'), 58^oC (30"), 72^oC (30")  × 10 cycles
                               94^oC (30"), 55^oC (30"), 72^oC (30")  × 25 cycles
                               72^oC (7')

Product size:
                                 HincII
                   687 bp --------------> 615 bp + 72 bp (in homozygote site present)

Reference: Unpublished, Kidd lab.
 
 

Kidd Lab Histograms for Allele Frequencies