GSM1689077: MC; Vitis vinifera; miRNA-Seq
The samples harvested at 0 h (none cold-treated, NCT) and 4 h (cold-treated, CT) were used to construct small RNA libraries. Total RNA was extracted from collected samples using Column Plant RNAOUT 2.0 kit (Tiandz, Beijing, China), following the instructions of the manufacturer. RNA concentration and quality were detected using Agilent 2100 Bioanalyzer. Total RNA was purified by electrophoretic separation on a 15% TBE urea denaturing polyacrylamide gel electrophoresis (PAGE) gel and small RNA regions corresponding to the 18–30 nt bands in the marker lane were excised and recovered. Then the 18–30 nt small RNAs were ligated to a 5’-adaptor and a 3’- adaptor sequentially by T4 RNA ligase (New England Biolabs, USA). The adapter-ligated small RNAs were subsequently transcribed into cDNA by SuperScript III Reverse Transcriptase (Invitrogen, USA) and then PCR-amplified for 17 cycles using the adaptor primers. The PCR products were purified for high-throughput sequencing.
GEO Accession
GSM1689077