Description
NGFR HincII Polymorphism

Procedure
HincII

(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 1.15 mM MgCl2, 50 mM KCl, 10 mM Tris HCl pH 8.4, 1M betane
NOTE THE BETANE

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: 94ºC, 5
94ºC (1'), 58ºC (30"), 72ºC (30") × 10 cycles
94ºC (30"), 58ºC (30"), 72ºC (30") 25 cycles
72ºC (7'),

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


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