GSM1531014: CK (without copper); Paeonia ostii; miRNA-Seq
For small RNA sequencing, the roots, stems and leaves of seedlings were separately harvested after 1, 6, 12 and 24 h of treatment and immediately frozen in liquid nitrogen for analysis.Total RNA was extracted from frozen roots, stems and leaves of Paeonia ostii with Trizol (Invitrogen, Carlsbad, CA, USA). Two sets of total RNA were prepared, with one derived from the original RNA pool prepared form Cu-treated tissues (roots, stems and leaves) (+Cu) at 1, 6, 12 and 24 h time points and the other form the RNA pool derived from Cu-free tissues (-Cu) at the same time points. The samples were then subjected to 15% denaturing polyacrylamide gel electrophoresis, after which the sRNA fragments of 18-28 nt were isolated from the gel and purified. Next, the sRNA molecules were ligated to a 5' adaptor and a 3' adaptor sequentially and then converted to DNA by RT-PCR. Finally, approximately 20 μg product of RT-PCR was sequenced directly using standard Illumina protocols.
GEO Accession
GSM1531014