Table 3

Dynamic range of E2, E6 and HMBS quantification by qPCR.

Template DNA per μl

Expected quantification in 2 μl

Observed quantification with percent change over expected




E2

E6

HMBS

E2/E6

E6 per diploid genome

E2

E6

HMBS

E2/E6

E6 per diploid genome












51 ng gDNA 10 pg pSP64 HPV16

760,000

760,000

28,571

1

53

893,063

+17.5%

616,115

-18.9%

49,816

+74.4%

1.4

25

11 ng gDNA 2.16 pg pSP64 HPV16

163,922

163,922

6,163

1

53

208,404

+27.1%

172,423

+5.2%

7,437

+20.7%

1.2

46

3 ng gDNA 60 fg pSP64 HPV16

44,665

44,665

1,679

1

53

26,956

-39.7%

40,597

-9.1%

1,047

+37.6%

0.7

78


Peripheral blood lymphocyte gDNA was spiked with a known quantity of HPV16 DNA (in the pSP64-HPV16 plasmid). The template DNA was serially diluted, and absolute amounts were measured by spectrophotometry. The template DNA values are given in the first column of the Table. The anticipated gene copy numbers for E2, E6 and HMBS were derived, and are shown in the 'Expected quantification' section. qPCR of the serially diluted samples was performed, and the observed gene copy numbers are reported in the 'Observed quantification' section.

Roberts et al. BMC Biotechnology 2008 8:57   doi:10.1186/1472-6750-8-57

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