Isoseq of Helianthus tuberosus: flower
High-quality total RNA was extracted from fresh root, quality checked by agarose gel electrophoresis and Nanodrop 2000 spectrophotometer, quantified by Qubit 2.0 and Agilent 2100 bioanalyzer. The mRNA was enriched, reverse transcribed into cDNA and amplified by NEBNext Single Cell / Low Input cDNA Synthesis & Amplification Module and Iso-Seq Express Oligo Kit. The amplified cDNA with peak length ~ 2 Kb was end-repaired, ligated, and purified into SMRT library by SMRTbell Express Template Prep Kit 2.0, and mixed with Sequel Sequencing Primer v4 and polymerase (Sequel II Binding Kit v2.1) before sequencing.