ena-STUDY-ETH ZURICH-29-09-2017-09:50:07:001-210
the transcriptome of a male Nepenthes khasiana inflorescence
Plants of Nepenthes khasiana (in vitro propagated material from Borneo Exotics (Pvt) Ltd., Sri Lanka) were grown in a greenhouse where they flowered regularly. For the transcriptome of a male inflorescence of N. khasiana (length c. 2 cm, many buds of c. 1-3 mm diameter each), we extracted RNA using the Total RNA Mini Kit (Plant) (Geneaid Biotech Ltd, New Taipei City, Taiwan) with the "PRB" lysis buffer, which yielded undegraded high quality RNA (RIN 7.1; Plant RNA Nano Assay, Agilent Bioanalyzer). A cDNA library was generated (NEBNext Ultra Directional RNA Library Prep Kit for Illumina, New England Biolabs, Ipswich MA, USA) and sequenced in one lane of the Illumina MiSeq for 150 bp paired-end reads (GDC ETHZ). A total of 18.7 million PE reads were obtained, and a reference transcriptome was de novo assembled by the Trinity pipeline (Grabherr et al., 2011). Best ORFs were extracted by the Transdecoder script.
Plants of Nepenthes khasiana (in vitro propagated material from Borneo Exotics (Pvt) Ltd., Sri Lanka) were grown in a greenhouse where they flowered regularly. For the transcriptome of a male inflorescence of N. khasiana (length c. 2 cm, many buds of c. 1-3 mm diameter each), we extracted RNA using the Total RNA Mini Kit (Plant) (Geneaid Biotech Ltd, New Taipei City, Taiwan) with the "PRB" lysis buffer, which yielded undegraded high quality RNA (RIN 7.1; Plant RNA Nano Assay, Agilent Bioanalyzer). A cDNA library was generated (NEBNext Ultra Directional RNA Library Prep Kit for Illumina, New England Biolabs, Ipswich MA, USA) and sequenced in one lane of the Illumina MiSeq for 150 bp paired-end reads (GDC ETHZ). A total of 18.7 million PE reads were obtained, and a reference transcriptome was de novo assembled by the Trinity pipeline (Grabherr et al., 2011). Best ORFs were extracted by the Transdecoder script.