transcriptome
A cDNA library was constructed using S. stapfianus mRNA as template to synthesize first-strand cDNA using a Modified Smart cDNA Synthesis Kit from Clontech Laboratories, Inc. (Takara Bio Company, Mountain View, CA, USA) according to the manufacturerOs instructions.   The primers used were: 5O-AAGCAGTGGTATCAACGCAGAGTGGCCATTACGGCC-GGG-3O for Smart IV, and 5O-TAGAGACCGAGGCGGCCGACATGTTTTGTTTTTT-TTTCTTTTTTTTTTVN-3O for modified CDS III/3O.  The second-strand cDNA was synthesized using the primer extension method (Clontech Laboratories, Inc.). Following gel electrophoresis of 5-mL samples, the remaining 95 mL of the second-strand product was applied to a Clontech TE-400 size-fractionation column (Clontech Laboratories, Inc.) as instructed by the manufacturer. The recovered product was then cleaned up (using Qiagen kit), eluted in 30 mL of nuclease-free water, and quantified by spectrophotometer (NanoDrop Technologies, Inc.). The cDNA was treated to generate blunt ends and ligated to 454-specific adaptors, then small fragments were removed using AMPureXP Beads (Agencourt Bioscience Corporation, Beverly, MA). The library was normalized to remove high-abundance cDNAs using the Trimmer kit (Evrogen, City, Russia). The cDNA was then quantified using a Qubit fluorometer (Invitrogen, Inc., City, CA, USA) and average fragment sizes were determined using a Bioanalyzer (Agilent, CA). 