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

BMC Bioinformatics

Table 1 RNaseP estimates including replicates

	predictions from basic model		predictions from expanded model
Initial number of amplicon molecules	40 cycles	60 cycles	40 cycles	60 cycles
1.25 × 10³	0.712 × 10³	0.866 × 10³	0.821 × 10³	1.20 × 10³
	0.734 × 10³	0.900 × 10³	0.831 × 10³	1.25 × 10³
	0.723 × 10³	0.888 × 10³	0.833 × 10³	1.20 × 10³
	0.734 × 10³	0.887 × 10³	0.830 × 10³	1.23 × 10³
2.5 × 10³	1.50 × 10³	1.8 × 10³	1.7 × 10³	2.4 × 10³
	1.5 × 10³	1.8 × 10³	1.7 × 10³	2.5e × 10³
	1.6 × 10³	2.0 × 10³	1.9 × 10³	2.7 × 10³
	1.7 × 10³	2.1 × 10³	2.0 × 10³	2.9 × 10³
5.0 × 10³	3.3 × 10³	3.9 × 10³	3.7 × 10³	5.1e × 10³
	3.2 × 10³	3.7 × 10³	3.6 × 10³	5.1e × 10³
	3.2 × 10³	3.7 × 10³	3.6 × 10³	5.3 × 10³
	3.0 × 10³	3.3 × 10³	3.5 × 10³	5.0 × 10³
1.0 × 10⁴	0.61 × 10⁴	0.66 × 10⁴	0.71 × 10⁴	1.2 × 10⁴
	0.60 × 10⁴	0.67 × 10⁴	0.83 × 10⁴	1.2e × 10⁴
	0.61 × 10⁴	0.67 × 10⁴	0.72 × 10⁴	1.2e × 10⁴
	0.61 × 10⁴	0.66 × 10⁴	0.71 × 10⁴	1.1 × 10⁴
2.0 × 10⁴	1.3 × 10⁴	1.3e × 10⁴	2.0 × 10⁴	2.4 × 10⁴
	1.3 × 10⁴	1.4 × 10⁴	1.5 × 10⁴	2.3 × 10⁴
	1.3 × 10⁴	1.5 × 10⁴	1.8 × 10⁴	2.4e × 10⁴
	1.4 × 10⁴	1.6 × 10⁴	2.2 × 10⁴	2.5 × 10⁴

Estimation results of initial amounts of RNaseP amplicon. Initial concentrations of probe and primer were 250 nmol/L and 900 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 additionally 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