Campanella_umbellaria transcriptome
Campanella umbellaria is environment samples collected in East Lake, Wuhan, China.  Each population was isolated in the laboratory using ophthalmological scissors and tweezers under the light microscope,and then picked out using dissecting needles and glass micropipettes under the light microscope  The RNA of isolated cells was stabilized using the RNA Protect Cell Reagent. Total RNA was extracted using the RNeasy Protect Cell Mini Kit according to protocol in TetraFGD. Total RNA concentrations were determined by the e-Spect ES-2 spectrophotometer and RNA integrity was verified using gel electrophoresis in denaturing 1.2% agarose.  RNA libraries were prepared for sequencing using standard Illumina protocols. Poly-A mRNAs were isolated using Sera-mag Magnetic Oligo (dT) beads from Illumina, and then fragmented using divalent cations at 94°C for 5 minutes. Double-stranded cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen, Camarillo, CA) with random hexamer primers from Illumina. The synthesized cDNA was subjected to end-repair, phosphorylation, 3' adenylation and adapter ligation in sequential. After these steps, cDNA fragments with an insert size of about 200bp were purified by gel electrophoresis. The purified cDNA template was enriched by PCR amplification and the quality of the RNA library was validated in a LightCycler480 (Roche Diagnostics) using an Illumina PhiX174 Control. Libraries were sequenced for 81bp at both ends (paired-end) using an Genome Analyzer IIX sequencer.