Absolute estimation of initial concentrations of amplicon in a real-time RT-PCR process

BMC Bioinformatics

Table 2 Cyp1B1 estimates with replicates

	predictions from basic model		predictions from expanded model
Initial number of amplicon molecules	40 cycles	60 cycles	40 cycles	60 cycles
1e6	0.59 × 10⁶	0.60 × 10⁶	0.66 × 10⁶	0.67 × 10⁶
	0.56 × 10⁶	0.58 × 10⁶	0.63 × 10⁶	0.63 × 10⁶
	0.56 × 10⁶	0.59 × 10⁶	0.65 × 10⁶	0.64 × 10⁶
1e5	0.48 × 10⁵	0.50 × 10⁵	0.55 × 10⁵	0.54 × 10⁵
	0.48 × 10⁵	0.52 × 10⁵	0.56 × 10⁵	0.58 × 10⁵
	0.53 × 10⁵	0.55 × 10⁵	0.61 × 10⁵	0.61 × 10⁵
1e4	0.46 × 10⁴	0.50 × 10⁴	0.47 × 10⁴	0.54 × 10⁴
	0.45 × 10⁴	0.49 × 10⁴	0.47 × 10⁴	0.53 × 10⁴
	0.50 × 10⁴	0.55 × 10⁴	0.52 × 10⁴	0.59 × 10⁴
1e3	0.52 × 10³	0.59 × 10³	0.57 × 10³	0.63 × 10³
	0.48 × 10³	0.53 × 10³	0.52 × 10³	0.58 × 10³
	0.51 × 10³	0.57 × 10³	0.57 × 10³	0.61 × 10³

Estimation results of initial amounts of Cyp1B1 amplicon. Initial concentrations of probe and primer were 100 nmol/L and 200 nmol/L respectively. The basic model accounts for the duplication of target sequence with release of fluorescence as well as re-annealing of previously formed DNA strands. The extended model also takes into account duplication of target sequence without probe attachment and hence without release of fluorescence. Results are given for individual replicates of each initial amount of amplicon.

ISSN: 1471-2105