biostudies-arrayexpressKarolina SobańskaMiscanthus sinensishttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15228Iso-Seq (PacBio) sequencing was performed to generate a reference transcriptome library of M. sinensis.biostudies-arrayexpressNucleic Acid Extraction - Frozen samples were homogenized using TissueLyser II (Qiagen, Hilden, Germany). RNA was extracted using an automated method, Maxwell® RSC Plant RNA Kit. The concentration and purity of RNA was measured using the Nanodrop 1000 spectrophotometer (ThermoFisher Scientific, Waltham, MA, USA). RNA integrity was determined using Bioanalyzer 2100 (Agilent, Santa Clara, CA, USA). Only RNA samples with 260/280 ratio higher than 1.8, 260/230 ratio higher than 2.0, and RIN (RNA integrity number) values higher than 8.0 were used for the RNA-seq analyses.Sample Collection - RNA was extracted from 2 cm middle fully expanded leaf segments. Tworeplicates per genotype were collected. Material was frozen in liquid nitrogen and stored at −80 °C. Frozen samples were homogenized using TissueLyser II (Qiagen, Hilden, Germany).Growth Protocol - Plants were first grown under greenhouse conditions (25°C day / 19°C night) and then subjected to either a constant control environment or a chilling treatment. Maternal plants were grown from 5 cm rhizome segments, each planted in 40 L containers filled with a 1:1 peat:sand mixture. A standardized nutrient solution (100 mg/kg N, 50 mg/kg P, 200 mg/kg K, 2 mg/kg Fe, 10 mg/kg Zn, 10 mg/kg Ca) with daily watering was implemented. Collected plantlets were initially cultivated in the greenhouse conditions under a 12-hour photoperiod using white fluorescent lighting with photosynthetically active radiation (PAR) at 750 μmol m⁻² s⁻¹. Day/night temperatures were held at 25/19°C, with 60% ± 10% of relative humidity. At the five-leaf stage, 20 individuals per genotype were relocated to a cultivation chamber at the Wielkopolska Centre for Advanced Technologies (Poznań, Poland). Light intensity, photoperiod and temperature were matched to the greenhouse setting to facilitate acclimation.Library Construction - Iso-Seq library preparation was done using PacBio full-length cDNA library preparation kit according to the manufacture’s protocol (Pacific Biosciences of California, Inc., Menlo Park, CA, USA) by the sequencing service provider.Sequencing - Iso-Seq sequencing was carried out on Sequel System.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - CCS sequences obtained in biological replications were processed (clustered, polished) by SMRT Link ver. 9.0 software to obtain full length transcript sequencesSequence Alignment - Row subread data were processed into consensus sequences by SMRT Link ver. 9.0 software according to SMRT Link Tools Reference guide ver. 9.0 separately for each biological replication to obtain replication-specific set of ccs.UnknownTranscriptomicsGenomicsProteomicsSequelRNA-seq of coding RNAMiscanthus sinensisKarolina SobańskaPaweł KrajewskifalseIso-Seq analysis of Miscanthus sinensis plantsIso-Seq (PacBio) sequencing was performed to generate a reference transcriptome library of M. sinensis.2025-11-21T00:00:00Z2025-11-21T11:26:10.609Z2025-06-13T12:32:46.151ZE-MTAB-15228ERP173449EFO_0002944EFO_0004170EFO_0003789EFO_0004917EFO_0005518EFO_0003816EFO_0003738EFO_0004184