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Table 8 Summarised results for the universal RNA standard spiked at the amplification step (part 2) during AuNP-interference assessment, using CFX Manager™ software

From: The presence of residual gold nanoparticles in samples interferes with the RT-qPCR assay used for gene expression profiling

  18S ACTB GAPDH GUSB HPRT1 HSP90 PPI SDH TBP YWHAZ
E (90 to 110%)a 116 to 124.7 84.7 to 106.8 84.2 to 98.2 108.8 to 117.5 96.2 to 100.9 99.1 to 106.4 105.3 to 154.1 135.0 to 145.9 105.6 to 364.8 90.0 to 98.5
R2 (> 0.980)b 0.994 to 1.000 0.961 to 0.998 0.995 to 1.000 0.986 to 1.000 0.990 to 1.000 0.995 to 1.000 0.970 to 1.000 0.956 to 0.994 0.945 to 0.997 0.989 to 1.000
Slope (− 3.1 to − 3.6)a − 2.845 to − 2.986  3.169 to − 3.751  3.365 to − 3.770 − 2.964 to  3.129  3.300 to  3.416  3.177 to  3.343 − 2.469 to  3.200 − 2.559 to − 2.695 − 1.499 to  3.195  3.358 to  3.588
NTC Cq 35.44 to 37.60 N/A N/A Peaks c 38.05 33.04 to 39.44 N/A 35.04 26.39 to 39.27 39.24
  1. Bold italic acceptable result; italics unacceptable result
  2. aFor an PCR efficiency (E) of 100%, the slope is − 3.32. A good reaction should have an efficiency between 90 and 110%, which corresponds to a slope between − 3.58 and − 3.10
  3. bThe linearity of the assay (R2), where R2 < 0.980 unacceptable; R2 ≥ 0.980 acceptable; R2 > 0.990 expected; R2 > 0.995 exceptional
  4. cThe peaks had shoulders, i.e. indication of secondary structures within the molecule