Illumina HiSeq 2000 paired-end Sequencing of KAS22022698 (KAE22247380)
Construction of reference genome of Castancea crenata
Total RNA was extracted using TRIzol reagent Invitrogen. Total NGS sequencing libraries were generated using TruSeq RNA Sample Prep Kit Illumina The cleaved mRNA fragments were reverse transcribed into first strand cDNA using random primers. Short fragments were purified with a QiaQuick PCR extraction kit and resolved with EB buffer for end reparation and addition of poly A. Subsequently, the short fragments were connected with sequencing adapters. Each library was separated by adjoining distinct MID tag. The resulting cDNA libraries were then paired-end sequence 2x101bp for individual samples with HiSeq??2000 system Illumina.