Pseudo-nitzschia multistriata
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.
ENA-FIRST-PUBLIC
2018-11-28T17:03:21Z
ENA-LAST-UPDATE
2018-08-09T10:07:26Z
External Id
SAMEA4823047
INSDC center name
Stazione Zoologica Anton Dohrn of Naples
INSDC first public
2018-11-28T17:03:21Z
INSDC last update
2018-08-09T10:07:26Z
INSDC status
public
Submitter Id
E-MTAB-7121:Sample 4
broker name
ArrayExpress
genotype
MRP3 overexpression
sample name
E-MTAB-7121:Sample 4
scientific_name
Pseudo-nitzschia multistriata
sex
mating type minus
strain
LV136T3