GSM5967850: Control rep 1; Oryza sativa; miRNA-Seq
Total RNA was isolated using modified guanidinium isothiocyanate (GITC) method (Chomzynski and Sacchi, 1987). Quantity and quality of RNA was assessed using a spectrophotometer (Bio-Rad, USA) followed by checking on MOPS-formaldehyde-agarose gel. Low molecular weight RNA fraction was extracted from the total RNA by resolving on a TBE-urea-PAGE gel and excising 15-30 nt portion. The Low molecular weight RNA was ligated with 5' and 3' RNA adapters using T4 RNA ligase. First strand cDNA was synthesized from the adapter ligated low molecular weight fraction and cDNA library was enriched using adapter-specific primers. The enriched libraries were purified, and the quality of the same was evaluated using Bioanalyzer (Agilent Technologies, USA). High-throughput sequencing of libraries was performed using Illumina GA IIx sequencing platform.