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Table 2 Detection yield, copy number and cycle threshold obtained in qPCR for the RT systems Sensiscript, Omniscript, and SuperScript II.

From: Detection limits of several commercial reverse transcriptase enzymes: impact on the low- and high-abundance transcript levels assessed by quantitative RT-PCR

Background RNA (ng)   0 10 25 50 100 1000 2000
RT systemsa Datab        
Sensiscript         
  DY (%) 0.62 1.36 7.69 14.57 16.64 13.61 10.63
Low copy copy num. ± SD 2 ± 2 3 ± 4 20 ± 18 37 ± 11 43 ± 21 35 ± 19 27 ± 24
  (Ct ± SD) (39.52 ± 0.68) (39.28 ± 0.71) (37.19 ± 2.45) (35.35 ± 0.43) (35.34 ± 0.72) (35.61 ± 0.93) (36.87 ± 2.71)
High copy DY (%) 3.92 25.61 29.79 39.26 38.77 48.33 50.41
  copy num. ± SD 10 068 ± 8880 65 700 ± 9128 76 433 ± 833 100 733 ± 18 034 99 467 ± 9 089 124 000 ± 21 071 129 333 ± 34 020
  (Ct ± SD) (30.6 ± 7.14) (24.24 ± 0.21) (24.02 ± 0.02) (23.62 ± 0.28) (23.64 ± 0.14) (23.29 ± 0.29) (23.67 ± 0.14)
GNPDA copy num. ± SD 0 ± 0 73 ± 25 236 ± 37 691 ± 39 2 150 ± 806 26 395 ± 1 404 42 488 ± 6 724
  (Ct ± SD) (40 ± 0) (32 ± 0.62) (30.03 ± 0.3) (28.21 ± 0.09) (26.42 ± 0.58) (22.22 ± 0.09) (21.45 ± 0.26)
Omniscript         
Low copy DY & copy num. N/A N/A N/A N/A N/A N/A N/A
  DY (%) 0.00 0.00 0.00 0.24 2.16 38.46 38.56
High copy copy num. ± SD 0 ± 0 0 ± 0 9 ± 15 623 ± 368 5 548 ± 4 152 98 664 ± 5 062 98 927 ± 11 165
  (Ct ± SD) (40 ± 0) (40 ± 0) (38.41 ± 2.75) (30.73 ± 1.59) (27.53 ± 2.1) (22.44 ± 0.08) (22.44 ± 0.17)
GNPDA copy num. ± %
(Ct ± SD)
primer dimer primer dimer primer dimer primer dimer primer dimer primer dimer primer dimer
SuperScript II         
  DY (%) 19.80 49.43 70.14 66.30 81.33 102.44 97.38
Low copy copy num. ± SD 51 ± 44 127 ± 39 180 ± 18 170 ± 49 209 ± 12 263 ± 40 250 ± 106
  (Ct ± SD) (34.05 ± 0.04) (33.41 ± 0.51) (32.72 ± 0.19) (32.82 ± 0.42) (32.47 ± 0.09) (32.15 ± 0.21) (32.32 ± 0.7)
  DY (%) 13.30 22.33 16.69 18.86 27.31 31.33 36.81
High copy copy num. ± SD 34 127 ± 4 284 57 277 ± 5 106 42 814 ± 26,175 48 389 ± 24 717 70 062 ± 8 760 80 366 ± 19 132 94 447 ± 10 237
  (Ct ± SD) (23.52 ± 0.19) (22.75 ± 0.14) (23.48 ± 1.19) (23.18 ± 0.98) (22.45 ± 0.19) (22.28 ± 0.4) (22 ± 0.16)
GNPDA copy num. ± SD 5 ± 9 102 ± 48 351 ± 201 1 122 ± 608 2 401 ± 752 19 088 ± 3 540 22 758 ± 1 815
  (Ct ± SD) (38.79 ± 2.1) (30.84 ± 1.17) (28.6 ± 1.12) (26.43 ± 0.91) (24.95 ± 0.57) (21.2 ± 0.35) (20.86 ± 0.14)
  1. aRT systems tested were Sensiscript. Omniscript. SuperScript II. SuperScript III. and PowerScript; low copy. RT samples spiked with 2.6 × 103 EGFP mRNA copies; high copy. RT samples spiked with 2.6 × 106 EGFP mRNA copies.
  2. bDY (detection yield) was calculated using the equation described in Results and the copy number (num.) was derived from the calibration curve for a given cycle threshold value (CT) [see Additional file 2];SD. standard deviation.