title: Equimolar pool of various sample sources and conditions (leaves and cell cultures)
organism: Quillaja brasiliensis
description: We collected seeds of Quillaja brasiliensis (A.St.-Hil. & Tul.) Mart. (Quillajaceae) from the city of Cangucu (Rio Grande do Sul, Brazil) in March/2018. After in vitro germination, we explanted the seedlings and submitted them to callogenesis or transferred them to the grow room at 25 C under a 16h/day photoperiod. We detached leaves from 2.8-year-old individuals for the experiments hereby presented. After leaf cleanup, we detached approximately 15 leaves and exposed them to ultraviolet radiation (UV-C germicide lamp, wavelength maximum 254 nm) or white light (control). After incubation, we froze the plant material in liquid nitrogen and stored it at -80 C until the next processing step. We obtained calluses from in vitro germinated explants and submitted them to cellular suspension induction. We maintained the cell cultures in MS medium supplemented with naphthaleneacetic acid (5 mg/L) and kinetin (0.1 mg/L) in the absence of light, using 250 mL culture flasks under the agitation of 120 RPM. After establishing the growth profile, we collected three flasks for each growth phase of the liquid cell cultures: lag phase (three days), log phase (seven days), and stationary phase (21 days). For the sample collection, cells were filtered using the Buchner funnel, washed with distilled water, and flash-frozen in liquid nitrogen. We stored the collected material at -80 C until the next processing step. We extracted RNA samples from different tissues and growth conditions using the cetyltrimethylammonium bromide method (CTAB) and performed their purification using the RNeasy MinElute Cleanup Kit (QIAGEN, Hilden, Germany). We quality-controlled the samples using fluorometric and spectrophotometric methods before submitting them to sequencing by Novogene (Beijing, People's Republic of China). We made an equimolar pool of the five sample types for sequencing using the Iso-Seq strategy (PacBio, Menlo Park, US) in circular consensus sequencing mode for the obtention of long reads.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-02-03
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-02-03
attributes.tag: External Id
attributes.value: SAMEA112328550
attributes.tag: INSDC center alias
attributes.value: IB-RC UNESP
attributes.tag: INSDC center name
attributes.value: ib-rc unesp
attributes.tag: INSDC first public
attributes.value: 2023-02-03T04:26:24Z
attributes.tag: INSDC last update
attributes.value: 2023-02-03T04:26:24Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: equimolar_pool_of_leaves_and_cell_cultures
attributes.tag: growth_phase
attributes.value: Multiple conditions
attributes.tag: sample name
attributes.value: equimolar_pool_of_leaves_and_cell_cultures
attributes.tag: scientific_name
attributes.value: Quillaja brasiliensis
attributes.tag: tissue_type
attributes.value: Leaf/Cell culture
attributes.tag: treatment
attributes.value: Multiple conditions