GSM4023430: sRNA apical spikelets; Oryza sativa; miRNA-Seq
Total RNA was extracted from the individual spikelet samples using Plant RNA purification reagent (Invitrogen). The quality of the RNA was checked for integrity on bioanalyzer, and only the RNA preparation with RNA integrity (RIN) greater than 7.0 was considered for the downstream application. Ion Total-Seq Kit v2.0 (Thermo Fisher) was used for enrichment of the sRNA population in the individual RNA preparation, ligation of barcoded adapters to sRNAs and conversion of the sRNAs to cDNAs. The cDNA libraries of sRNAs prepared was verified for size distribution using Agilent 2100 Bioanalyzer. The individual libraries were diluted to 100 pM and pooled in equimolar concentrations for clonal amplification and sequencing. The clonal amplification was done on ion sphere particles (ISs) by emulsion PCR. ISPs positive templates were loaded on Ion PI chip kit (Thermo Fisher) after denaturing and sequencing was on the Ion Proton System
GEO Accession
GSM4023430