vg gDNA quantification

qRT-PCR results showed vg mRNA expressions were significantly higher in body walls of winged aphids relative to wingless aphids at the third nymphs. A. pisum genome shows a large number of gene duplications [55]. So, we determined if gene DNA copy number contributes to the difference by using qRT-PCR: the genomic DNA was isolated from body wall of 20 third instar of wing morphs using DNAzol (MRC) according to the manufacturer’s instructions. qRT-PCR were performed as described above, except that the primers were designed based on the vg exon sequences which were confirmed by aligning vg ORF nucleotide sequences with A. pisum genome in NCBI.