Table 3

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

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)


rxn, reaction.

ainvA mRNA copies per reaction are mean values of 6 determinations ± standard deviations.

binvA DNA copies per reaction are mean values of 6 determinations ± standard deviations.

c- If <4 is negative or inconclusive for invA mRNA presence due to DNA contamination.

Detection limit of the invA qRT-PCR and invA qPCR are 40 and 10 copies per reaction, respectively [7].

Rump et al. BMC Research Notes 2010 3:211   doi:10.1186/1756-0500-3-211

Open Data