Saturday, May 15, 2004

Description
PLAT Alu Polymorphism

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

Primer 1: PLAT.A: 5'-GTG AAA AGC AAG GTC TAC CAG-3 100 ng/Rx
Primer 2: PLAT.B: 5'-GAC ACC GAG TTC ATC TTG AC-3' 100 ng/Rx

dNTPs: Final Concentration 200 uM each

Buffer: Final Concentration 50 mM KCl 10 mM Tris-HCl (pH8.4) 2.5 mM MgCl2

Taq DNA Polymerase: 0.5 U/reaction
(Perkin Elmer Amplitaq DNA polymoerase 5 U/ul, Cat. No. N808-0145)

PCR Cycling Profile: 95oC, 5'
95oC (1'), 60oC (1'), 72oC (1') x 30

Product size: 570 bp and 260 bp for heterozygous (+/-) sample

Friezner-Degen, SJ, B Rajput, E. Reich, 1986. The human tissue plasminogen activator gene. J. Biol. Chem 261:6972-6985.

Tishkoff, SA, Ruano G, Kidd JR, Kidd KK, 1996. Distribution and frequency of a polymorphic Alu insertion at the plasminogen activator locus in humans. Hum Genet 97:759-764.


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