Incorporation or lack of incorporation of deoxynucleotide triphosphates
(dNTPs) indicates the DNA sequence.
Optimal PCR conditions for triplex amplification of factor V Leiden, factor II G20210A, and MTHFR C677T were as follows: 10 ng of pure genomic DNA, 10 pmol of each primer, 200 [micro]M each deoxynucleotide triphosphate
(dNTP), 3 mM Mg[Cl.
2], and 2 nmol of each deoxynucleotide triphosphate
2 [micro]M each primer, 200 [micro]M each deoxynucleotide triphosphate
, 1x Q solution (Qiagen), and 1 U of HotStarTaq DNA polymerase in 1x supplied PCR buffer (Qiagen).
025 U/[micro]L AmpliTaq Gold DNA polymerase, 200 [micro]M each deoxynucleotide triphosphate
, 10 mM Tris-HCl (pH 8.
8 x Post-extension 72 5 5 1 PCR reactions were performed in 25 PL containing 50 ng of DNA, 100 [micro]M each deoxynucleotide triphosphate
with 100 [micro]mol/L each deoxynucleotide triphosphate