Assessment and validation of a suite of reverse transcription-quantitative PCR reference genes for analyses of density-dependent behavioural plasticity in the Australian plague locust

Table 1 Primer sequences, amplicon lengths and reaction efficiencies in RT-qPCR study.

Symbol	Primer sequence (5'-3')	size (bp)	Tm (°C)	Primer	E (error)	SD(E)
18SrRNA	F: CTGAGAAACGGCTACCACATC R: ACCAGACTTGCCCTCCAAT	171	84.9	0.25	1.91 (0.02)	0.02
Arm	F: ACTTCTTATGAGAGCATTCCAGGAT R: CCTTCAACAATTTCTTCCATGC	114	83.2	0.3	1.81 (0.03)	0.01
EF1a	F: AGCCCAGGAGATGGGTAAAG R: CTCTGTGGCCTGGAGCATC	155	81.4	0.3	1.99 (0.08)	0.04
RpL32	F: ACTGGAAGTCTTGATGATGCAG R: CTGAGCCCGTTCTACAATAGC	97	78.6	0.25	1.97 (0.05)	0.04
GAPDH	F: AATTGCCTGGCACCATTG R: CGCCACAACTTTCCAGATG	128	80.7	0.3	1.95 (0.06)	0.00
Actin	F: TTGTGTTGGATTCTGGTGATG R: GAAGCTGTAGCCCCTCTCAG	149	83.5	0.5	1.66 (0.01)	0.02
SDHa	F: CCACTGAAACTGATCCAAGAGAG R: TCCTGCTCCATTAACTAAGCAAC	98	76.2	0.3	1.90 (0.05)	0.10
AnnIX	F: GGAACTGATGAGGAAGCCATT R: TGGCCTGAAGTGTCTCCTTT	134	77.2	0.5	1.91 (0.04)	0.05

size: size of the cDNA amplicon; Tm: melt temperature of the PCR amplicon averaged over the 15 biological samples and 3 technical replicates; Primer: primer concentration in μM; E: in-run PCR reaction efficiency (fold increase per cycle) computed from the standard curve of a serial dilution included in the same plate as for samples S1 to S15 and using the formulae 10^{-(1/-slope of the standard curve)}; error: the mean square error of the standard curve; SD(E): standard deviation of the PCR efficiencies estimated from duplicated standard curves (different RT-qPCR runs and cDNA templates).

ISSN: 1471-2199