Triticum aestivum Chinese Spring
Plants were grown in greenhouse conditions under natural day light supplemented with artificial source of light. The total RNA was isolated from 4 week seedlings using the RNeasy Plant Mini Kit (QIAGEN). First-Strand cDNA synthesis was performed according SMART cDNA synthesis technology (Clontech Laboratories, Inc.) using 3' SMART CDS modified Primer II A primer (5'-AAG CAG TGG TAT CAA CGC AGA GTA CTT TTG T(9) C T(10) VN-3') and SuperScript III reverse transcriptase (Invitrogen).   Double-stranded cDNA was amplified by long-distance (LD) PCR using Advantage 2 PCR Enzyme System (Clontech Laboratories, Inc).   The double-stranded cDNA was normalized using TRIMMER cDNA normalization Kit (EVROGEN), which is based on a unique DSN (duplex-specific nuclease) normalization technology and is specially developed for normalization of cDNA enriched with full-length sequences.