RNA extraction, cDNA synthesis and RT-PCR
Total RNA was extracted with RLT buffer and purified using an RNeasy
mini kit (Qiagen®) according to manufacturer’s instructions. RNA yield
was determined using a NanoDrop ND-8000 spectrometer. Genomic DNA was
removed by DNase I incubation using the RNase-Free DNase Set (Qiagen®)
and RNA was converted to cDNA using SensiFAST cDNA synthesis kit
(Geneflow). Quantitative PCR (qPCR) was completed using the SYBR® 3
Green master mix, with amplification carried out in triplicate using a
Rotor-Gene® Q (Qiagen®). Expression of target genes were normalised to
reference gene 36B4. Primer sequences are listed in Table S1.