Table 2 |
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|
Inability of RT-qPCR to detect invA mRNA in RNA extracts not treated with DNase I [] |
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|
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 |
|
|
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|
RiboPure-Bacteria |
Ambion |
(1.9 ± 1.6) × 106 [25.5 ± 1.8] |
(1.8 ± 1.8) × 106 [24.8 ± 1.4] |
-0. 7 (Negative) |
|
PureLink RNA Mini |
Invitrogen |
(1.2 ± 0.5) × 108 [18.8 ± 0.6] |
(1.5 ± 0.8) × 108 [18.5 ± 0.7] |
-0.3 (Negative) |
|
RNeasy Mini |
QIAGEN |
(7.9 ± 2.2) × 107 [19.3 ± 0.4] |
(1.1 ± 0.3) × 108 [18.9 ± 0.4] |
-0.4 (Negative) |
|
MasterPure RNA Purification |
EPICENTRE Biotechnologies |
(5.8 ± 0.8) × 108 [16.4 ± 0.2] |
(8.8 ± 0.9) × 108 [15.9 ± 0.2] |
-0.3 (Negative) |
|
UltraClean Microbial RNA Isolation |
MoBio |
(2.2 ± 0.6) × 108 [17.8 ± 0.4] |
(3.4 ± 0.8) × 108 [17.3 ± 0.3] |
-0.5 (Negative) |
|
|
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|
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] |
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|
Rump et al. BMC Research Notes 2010 3:211 doi:10.1186/1756-0500-3-211 |
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