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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].
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BMC Research Notes

ISSN: 1756-0500

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