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 2 Details of PCR amplification of a genomic DNA template.

Symbol	size (bp)	Tm (°C)	Cq (cycles)
18SrRNA	~170	84.3	14.8
Arm	no amplification	no amplification	no amplification
EF1a ^#	light smear	3 peaks: 76.6; 79.8; 83.7	35.8
RpL32	~100	78.4	28.8
GAPDH ^#	light smear	79.8	34.9
Actin	~650	no amplification	no amplification
SDHa	no amplification	no amplification	no amplification
AnnIX	light smear	no amplification	no amplification

Genomic DNA amplicons were sized on an agarose gel following 30 cycles of PCR amplification using a Taq polymerase from Qiagen, a melting temperature of 58°C, and 50 ng of template. The melting temperatures (Tm) and Cq values were determined by quantitative PCR on a LightCycler 480 Instrument (Roche) as detailed in the main text and on 5 ng of genomic DNA. ^# means that primers span an exon-exon boundary. Alignments of complementary and genomic DNA sequences can be found for Actin, GAPDH, and EF1a in Additional file 3.

ISSN: 1471-2199