. Author manuscript; available in PMC: 2007 Sep 1.

Published in final edited form as: Blood Cells Mol Dis. 2006 Aug 2;37(2):100–106. doi: 10.1016/j.bcmd.2006.06.003

Table 1.

Primer sequence, PCR conditions, primer mix and reaction mixture specifications

	Specifications
Primers	Primer FVL(G1961A)-C 5’-tgt tat cac act ggt gct taa-3’ Primer FVL(G1961A)-N 5’-cag atc cct gga cag acg-3’ Primer FVL(G1961A)-M 5’- cag atc cct gga cag aca-3’ Primer Factor IX-F 5’-ctc ctg cag cat tga ggg aga tgg aca tt-3’ Primer Factor IX-R 5’-ctc gaa ttc ggc aag cat atc aat gta t-3’
Primer Mix	Factor IX-F@10μM (0.10μM) Factor IX-R@10μM (0.10μM) FVLM (G1961A)-C@10μM (1.20μM), FVL (G1961A)-N@10μM (1.20μM) FVL (G1961A)-M@10μM (1.20μM) sterile water (qs)
Reaction mixture	10X Thermopol Buffer II (2X), 100mM MgSO₄ (3.3mM) 10mM dNTP solution^*(100μM) Betaine (20%) Taq Polymerase (0.117U/μl) Sterile water
PCR conditions	94ºC x 5’ / (94ºC x 30”; 55ºC x 30”; 72ºC x 15”) x 10 / (94ºC x 30”; 50ºC x 30”; 72ºC x 15”) x 20 / 72ºC x 5’ / 4ºC Pause. Heated lid at 98ºC.

10mM dNTP solution is composed of 2.5mM each of dATP, dCTP, dGTP and dTTP