biostudies-arrayexpressMetabolomicsUnknownTranscriptomicsGenomicsProteomicsMagdalena KrocSequel IIRNA-seq of coding RNALupinus angustifoliusLupinus angustifoliushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-16429The aim of this experiment was to obtain full length transcript sequences of genes expressed in leaves and stems of narrow-leafed lupin (Lupinus angustifolius L.) using PacBio Iso-Seq. Total RNA was isolated from leaves and stems of five narrow-leafed lupin (Lupinus angustifolius L.) genotypes with contrasting alkaloid regulation, including an iucundus low-alkaloid line (83A:476), a bitter Iucundus line (P27255) and three Bryansk low-alkaloid lines carrying the Iucundus-type RAP2-7 allele: 95826 (Bryanskij-35), 95927 (Bryanskij-123) and 95928 (Bryanskij-237/83). Plants, in five replicates per genotype, were grown under controlled conditions optimal for growth, and leaf and stem samples were collected at flowering stage. For each genotype and organ, total RNA from five biological replicates was pooled in equal amounts and used for PacBio Iso-Seq library preparation and sequencing.biostudies-arrayexpressGrowth Protocol - Plants were cultivated in a growth chamber (16 h photoperiod, 22/18 °C day/night, 55–60% relative humidity, until they reached whole plant flowering stage.Sample Collection - Fully expanded young leaves from plants that had reached the whole plant flowering stage were collected into sterile plastic tubes, immediately flash frozen in liquid nitrogen and stored at −80 °C until nucleic acid extraction. Immediately after leaf sampling, stem tissue was collected from the same plants by excising segments from the upper, middle and lower parts of the main stem. Stem segments were placed into sterile microcentrifuge tubes, flash frozen in liquid nitrogen and stored at −80 °C until RNA isolation.Nucleic Acid Extraction - Total RNA was isolated from 30 mg of frozen, ground organ tissue using a Maxwell RSC Plant RNA Kit (Promega, Madison, WI, USA). RNA concentration and integrity were assessed with a NanoDrop spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) and an Agilent 2100 Bioanalyzer (Agilent Technologies, Waldbronn, Germany), with a minimum RNA integrity number of 8.Sequencing - Sequencing was performed on the PacBio Sequel II system (HiFi/CCS mode; ≥20 GB raw data per sample) at Novogene Co. Ltd. Circular consensus sequences were generated using pbccs version 6.4.0 (minimum predicted accuracy 0.9).Library Construction - Total RNA from five plants per accession was pooled in equal amounts to prepare a single Iso-Seq library. Libraries were constructed following PacBio’s Iso-Seq sequencing protocol combining non-size selected and size selected fractions. The first strand cDNA was synthesized using the SMARTer PCR cDNA Synthesis Kit (Clontech, CA, USA), amplified, and processed with the SMRTbell Template Prep Kit 1.0 (Pacific Biosciences, CA, USA).OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesKatarzyna CzepielMagdalena KrocGrzegorz KoczykfalsePacBio Iso-seq of Lupinus angustifolius L. leaf and stem transcriptomesThe aim of this experiment was to obtain full length transcript sequences of genes expressed in leaves and stems of narrow-leafed lupin (Lupinus angustifolius L.) using PacBio Iso-Seq. Total RNA was isolated from leaves and stems of five narrow-leafed lupin (Lupinus angustifolius L.) genotypes with contrasting alkaloid regulation, including an iucundus low-alkaloid line (83A:476), a bitter Iucundus line (P27255) and three Bryansk low-alkaloid lines carrying the Iucundus-type RAP2-7 allele: 95826 (Bryanskij-35), 95927 (Bryanskij-123) and 95928 (Bryanskij-237/83). Plants, in five replicates per genotype, were grown under controlled conditions optimal for growth, and leaf and stem samples were collected at flowering stage. For each genotype and organ, total RNA from five biological replicates was pooled in equal amounts and used for PacBio Iso-Seq library preparation and sequencing.2026-06-30T00:00:00Z2026-06-30T01:00:54.362Z2025-12-19T12:32:43.936ZE-MTAB-16429ERP186831EFO_0002944EFO_0004170EFO_0003789EFO_0005518EFO_0003738EFO_0004184