title: Transcriptomes of Leymus chinensis under saline-alkaline treatment using 454 pyrosequencing
design: The quality of two total RNAs was checked using the NanoDrop Spectrometer (ND-1000 Spectrophotometer, Peqlab). The mRNAs were isolated from total RNAs using the PolyATtract® mRNA Isolation Systems kit (Promega, company) and condensed using Reasy RNA cleaning kit (QIAGEN, Germany), their concentration and purity were determined using the Agilent 2100 Bioanalyzer (RNA Nano Chip, Agilent). Using RNA Fragment reagent kit (Illumina, company) and Reasy RNA cleaning kit (QIAGEN, Germany) to fragment and retrieve the mRNA that has been condensed for 1min. Then using random primer and MMLV to synthesize the first chain, and using DNA Polymerase I and RNase H to synthesize the second chain. Finally using Reasy RNA cleaning kit (QIAGEN, Germany) to retrieve cDNA, and using the Agilent 2100 Bioanalyzer to check the quality of cDNA. All procedures were applied according to the manufacturer’s protocol.