Hiseq technique

We performed whole-exome sequencing (WES) using the Illumina HiSeq technique of next-generation sequencing. We create the exome-wide libraries from DNA using the SureSelect AllExome V4 50Mb from Agilent (Santa Clara, CA, USA). Then, we generated paired-end sequence data using Illumina HiSeq machines (Illumina, Inc., San Diego, CA, USA) (8plex sequencing).

Fifteen libraries were constructed from triplicated floral buds randomly selected at the five dormancy stages for transcriptome sequencing. The total RNA extraction was performed by following the protocol of TRIzol (TaKaRa, Dalian, China), and the extracted RNA was treated with DNase-I, Oligo (dT) to obtain mRNA. The quality assessment of extracted RNA regarding RNA integrity and contamination was estimated with the help of the Agilent 2100 Bioanalyzer system (Agilent Technologies, CA, United States) and 1% agarose gel electrophoresis, respectively. For the construction of paired-end libraries, 3 μg of RNA extracted from each sample was utilized and put in fragmentation buffer, followed by the preparation of cDNA from mRNA. The quantification and qualification of the sample library were assessed with the Agilent 2100 Bioanalyzer and the ABI StepOnePlus Real-Time System.
Moreover, according to the instructions recommended by the manufacturer (Illumina^®), libraries were created by utilizing Kit. For RNA-Seq, the Illumina HiSeq technique was used. Furthermore, the paired-end libraries (read length: 150 bp) were sequenced using the Illumina HiSeq 2500 platform.