Strand-specific RNA-seq libraries were constructed  and sequenced on a HiSeq2000 system
Patches of 5cm×5cm×5cm grass lawn was planted in pots (10cm) in a greenhouse with 14h light period. For cold treatment, the grass pots were transferred to a growth chamber with 4°C constant temperature. Pots in room temperature were used as control (CK). During the cold treatment, light period is14h, and light intensity is 100 µmol m-2 s-1. After 2h and 72h of treatment, the second leaf from the top was collected and then frozen with liquid nitrogen and stored in a -80°C refrigerator.  Total RNA was extracted using the Universal Plant Total RNA Extraction Kit RP3301 (Produced by Bio Take Corporation) following the manufacturer’s instruction. RAN integrity was confirmed using Agilent 2100 BioAnalyzer. Samples with RIN=7 and 28S:18S ratio=1.5:1, total amount 0.1?4ug were used in for the following research. Strand-specific RNA-seq libraries were constructed and sequenced on a HiSeq2000 system according to the manufacturer’s instructions in the Capitalbio company .
Minzhu University of china
CK