GSM1303739: Reactivating  vascular cambium; Cunninghamia lanceolata; RNA-Seq
Total RNAs of dormant, reactivating, and active cambial meristem, respectively were isolated using the Concert Plant RNA Reagent. 16-30 nt sRNAs were purified and two adaptors were sequentially ligated to the 5' and 3' ends of purified sRNAs. To separate the small RNA (16-30 nt) from total RNA: 1) Mix total RNA with 2X loading Buffer of equal volume and then incubate it at 65C for 5 minutes to break the second structure within RNA. 2) After pre-running of the 15% TBU PAGE for 15 minutes, load the samples, 10bp DNA ladder and 14-30nt RNA ladder into the gel separately. Run the gel at 180V for 1 hour. 3) Cut out the band of gel corresponding to the 16-30nt in the marker lane. Adapter sequences: 5' RNA Adapter: 5' GUUCAGAGUUCUACAGUCCGACGAUC 3' RNA Adapter: 5' P-UCGUAUGCCGUCUUCUGCUUGUidT The ligation products were further purified and then RT-PCR amplified. The amplification products were purified and utilized for sequencing on an Illumina 2G Genome Analyzer at the Beijing Genomics Institute, Shenzhen.
GEO Accession
GSM1303739