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Table 4 Summarised results of the universal RNA standard spiked before transcription (part 1), obtained 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

98.9 to 126.3%

99.7 to 158.7%

82.2 to 89.1%

102.7 to 105.7%

91.3 to 94.7%

100 to 101.5%

104 to 127.7%

125.2 to 146.6%

99.7 to 101.2%

81.7 to 88.2%

R2 (> 0.980)b

0.973 to 0.996

0.834 to 0.996

0.955 to 0.997

0.960 to 0.993

0.981 to 0.999

0.987 to 0.997

0.969 to 0.992

0.910 to 0.996  

0.992 to 0.997

0.984  to 0.995

Slope (− 3.1 to − 3.6)a

− 2.819 to − 3.348

− 2.422 to − 3.330

 3.614 to − 3.842

 3.193 to  3.258

 3.457 to  3.507

 3.286 to  3.321

− 2.798 to −3.226 

− 2.551 to − 2.837

 3.236 to  3.329

− 3.642 to − 3.855

NTC Cq

35.85 to 38

N/A

N/A

N/A

N/A

N/A

N/A

N/A

N/A

N/A

  1. Bold italics acceptable result; italics unacceptable result
  2. aFor a 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