Skip to main content

Table 3 Effect of DNase I treatment of RNA extracts on ability to detect invA mRNA by RT-qPCR

From: Comparison of commercial RNA extraction kits for preparation of DNA-free total RNA from Salmonella cells

RNA extraction kit Manufacturer invA mRNA copies/rxn
RT-qPCR [Cq]a
invA DNA copies/rxn
qPCR [Cq]b
invA mRNA presence
(Cq qPCR - Cq RT-qPCR)c
RiboPure-Bacteria Ambion (5.46 ± 0.88) × 105 [26.5 ± 0.9] <10 [37.1 ± 1.5] 10.6 (Positive)
PureLink RNA Mini Invitrogen (7.71 ± 1.89) × 105 [26.0 ± 1.2] <10 [37.5 ± 1.8] 11.5 (Positive)
RNeasy Mini QIAGEN (7.82 ± 1.25) × 106 [21.5 ± 0.5] <10 [36.5 ± 1.7] 15 (Positive)
MasterPure RNA Purification EPICENTRE Biotechnologies (1.19 ± 2.38) × 108 [18.7 ± 1.0] (3.10 ± 2.60) × 107 [21.5 ± 1.2] 2.8 (Negative)
UltraClean Microbial RNA Isolation MoBio (1.28 ± 1.12) × 105 [28.6 ± 0.6] (1.00 ± 1.20) × 103 [33.2 ± 1.5] 4.6 (Positive)
  1. rxn, reaction.
  2. ainvA mRNA copies per reaction are mean values of 6 determinations ± standard deviations.
  3. binvA DNA copies per reaction are mean values of 6 determinations ± standard deviations.
  4. c- If <4 is negative or inconclusive for invA mRNA presence due to DNA contamination.
  5. Detection limit of the invA qRT-PCR and invA qPCR are 40 and 10 copies per reaction, respectively [7].