GSM2011601: XSCD1dgd; Glycine max; miRNA-Seq
Leaves of three replicates with 24h chilling treatment and control were collected, respectively, and stored at -80 °C. For Solexa sequencing, three replicates were pooled together from chilling treatment and control, respectively. Total RNA was extracted separately with TRIzol reagent (Invitrogen, USA) according to the manufacturer’s instructions. RQ1 RNase-Free DNase (Promega, USA) was used to remove genomic DNA contamination. The total RNA quantity and purity were analysis by Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit (Agilent, CA, USA). About 1 μg of total RNA were used to construct small RNA library following the protocol of TruSeq Small RNA Sample Prep Kits (Illumina, San Diego, USA). Then we performed the single-end sequencing on an Illumina Hiseq2500 at the LC-BIO (Hangzhou, China) following a recommended protocol 45.A degradome li-brary was constructed from vegetable soybean leaves to predict the potential target mRNAs following the methods described previously by German et al.48. The purified cDNA li-brary was used for cluster genera-tion on Il-lumina’s Cluster Station and then sequenced on Il-lumina Hiseq 2500 following vendor’s in-struction for running the in-strument. Raw sequencing reads were obtained using Il-lumina’s Pipeline v1.5 software.