Comav cucurbita MU16
Total RNA was extracted from each tissue using the TRIzol Reagent (Invitrogen, USA). mRNA was purified from the total RNA using the illustraTM mRNA Purification Kit (GE Healthcare, Amersham Bioscience, Buckinghamshire, UK). Double-stranded cDNA was then synthesized from the RNA pool with the SMART cDNA Library Construction Kit (Clontech, USA). A normalization step was carried out with TRIMMER Kit (Evrogen, Moscow, Russia) in order to prevent over-representation of the most common transcripts. The PCR products of cDNA were purified using the QIAquick PCR Purification Kit (QIAGEN, Germany). Normalization quality of cDNAs libraries was checked by quantitative PCR.