title: Sample 2
organism: Pseudo-nitzschia multistriata
description: Protocols: Wild type and transgenic cultures, grown at 18°C with 12:12 D/L, were collected during the late exponential phase of growth, in the light phase. The MRP3 CDS amplified from the cDNA of the MT+ strain LV149 was cloned downstream of the P. multistriata histone H4 promoter, producing the overexpression vector PmH4pMRP3At. This vector was transformed in the MT- strain LV136 using biolistic transformation, with the protocol described in Sabatino et al., 2015. The transformed strain which had incorporated the transgene was named LV136T3. RNA was extracted following the manufacturer's instructions (Trizol reagent, Invitrogen) and the genomic DNA contamination was removed by DNase I treatment (RNase-Free DNase Set Qiagen) followed by RNA purification using RNeasy Plant Mini Kit (Qiagen). The quantity of RNA was determined by Qubit assay (Qubit® 2.0 Fluorometer, Life Technologies) and integrity was assessed by Bioanalyzer (2100 Bioanalyzer Instruments, Agilent Technologies) as well as by running samples on a 1.5% agarose gel. Transcriptome sequencing was conducted using the Ion P1 sequencing Kit v2 (Thermo Fisher Scientific) on an Ion Proton™ sequencer, following Manufacturer's instructions. Samples were run on two Ion P1 Chip Kit v3 chips (Thermo Fisher Scientific). All the libraries were sequenced in both chips to avoid the influence of batch effects.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2018-11-28T17:03:21Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-08-09T10:07:26Z
attributes.tag: External Id
attributes.value: SAMEA4823045
attributes.tag: INSDC center name
attributes.value: Stazione Zoologica Anton Dohrn of Naples
attributes.tag: INSDC first public
attributes.value: 2018-11-28T17:03:21Z
attributes.tag: INSDC last update
attributes.value: 2018-08-09T10:07:26Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7121:Sample 2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: sample name
attributes.value: E-MTAB-7121:Sample 2
attributes.tag: scientific_name
attributes.value: Pseudo-nitzschia multistriata
attributes.tag: sex
attributes.value: mating type minus
attributes.tag: strain
attributes.value: LV136