RNA-seq of Mitragyna speciosa: young leaf
Total RNA was extracted from young leaves using RNeasy Plant Mini Kit (Qiagen, USA) and from mature leaves using InnuPrep Plant RNA Kit (Analytik Jena, Germany). Three biological replicates from each young and mature group with the highest RIN score were chosen for RNA sequencing. From each sample, 1 g of total RNA were used for cDNA library construction by using Agilent SureSelect Strand-Specific RNA Library kit for Illumina Multiplexed Sequencing (mRNA Library Preparation Protocol). The cDNA poly (A)-containing mRNA libraries were then sequenced using Illumina HiSeq 4000 platform at Theragene Etex Bio Institute (Gyeonggi-do, Republic of Korea).