title: GSM2641680: Small RNAs in Hibiscus syriacus (Leaf); Hibiscus syriacus; miRNA-Seq
library_construction_protocol: Total RNAs were extracted from Hibiscus syriacus tissue samples (leaf, root, flower, and ovary) using TRI Reagent solution (MRC), according to the manufacturer’s protocol. Both the flower and ovary were pretreated with CTAB buffer [2% CTAB, 2% polyvinylpyrrolidone K 30, 100 mM Tris (pH 8.0), 25 mM EDTA, 2 M NaCl, and 2% β-mercaptoethanol] followed by TRI Reagent extraction, as previously described (Peng et al., 2014). A small RNA sequencing library was constructed using the Small RNA Sample Prep Kit (Illumina), according to the manufacturer’s instructions. As previously described (Hwang et al., 2013), 10 μg of total RNAs was resolved using 15% urea-PAGE, and size fractions of small RNAs between 18 and 30 nt were prepared. The prepared small RNAs were ligated to the 3′ and 5′ adapter, and cDNA was synthesized using RT-PCR. cDNAs were amplified using PCR and submitted for Illumina/Solexa sequencing.
attributes.tag: GEO Accession
attributes.value: GSM2641680