Saturday, May 15, 2004

Description
NGFR HindIII Polymorphism

Procedure
HindIII

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

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

Primer 1. NGFRHND3F 5' CCC CTT GCGG ATG TTC TC 3' 90ng/reaction
Primer 2. NGFRHND3R 5' GTG GGA TGG CTG GAC CTG 3' 90 ng/reaction

dNTPs: Final Concentration 200 M 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/l, Cat. No. N808-0145)

PCR Cycling Profile: 94C, 5 min
94C (30"), 58C (30"), 72C (30") x 30 cycles
72C (7')

Product size: HindIII
492------------------> 324 and 168 bp in heterozygote


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