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The first nucleotide dispensed, dATP, is not incorporated into either nascent strand, whereas the next one, dGTP, is incorporated twice on the strand for the wild-type allele and only once on the strand for the mutant allele, causing the polymerase molecules to go out of phase.
1 mmol/L each of 7-deaza-2'-dGTP (7-deaza-dGTP) and dGTP (Roche Molecular Diagnostics).
2] (Invitrogen, USA), 200 [micro]M of dNTP (Invitrogen, USA) containing each of dATP, dCTP, dGTP, dTTP the PCR primers, 2.
2]; 100 [micro]M each of dATP, dCTP, dGTP, and dTTP; 0.
5 [micro]l dNTP mix (10 mM each of dATP, dCTP, dGTP and dTTP), 1.
2]; 200 [micro]M (each) dATP, dCTP, dGTP, and dTTP; 1.
1 [mu]l of the cDNA obtained from the reverse transcription reactions was amplified in a total volume of 20 [mu]l consisting of 1 * GoTaq reaction buffer, 2U GoTaq DNA polymerase, 200 nM each of dATP, dCTP, dGTP, and dUTP, gene-specific primers that were added at a final concentration of 200 nM (all reagents were from Promega, USA).
5 mM each of dATP, dCTP, dGTP and dTTP), 1 of 10 pM of each primer and 1 U HS Prime Taq (GeNet Bio, Korea) in a 25 | l reaction volume.
2]; 200 [micro]M (each) deoxynucleoside triphosphates dATP, dCTP, dGTP, and dUTP; and 0.
2 mmol/L each of dATP, dGTP, dTTP, and dCTP; 8 pmol of each primer; 0.
The PCR mixture contained 1 x PCR buffer; dATP,dCTP, dGTP, dTTP (2 [micro]M each); primers (100 nM); Mg[Cl.
It is believed that high concentrations of dGTP cause an imbalance in the intra-cellular deoxynucleotide, ultimately resulting in the cell death of certain cancers.
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