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Sample GSM4769700 Query DataSets for GSM4769700
Status Public on Feb 24, 2021
Title 9RNA15.3
Sample type SRA
 
Source name leaves
Organism Nicotiana tabacum
Characteristics cultivar: Xanthi NN
transgenic line: Dclcyb1 transgenic line L15
replicate: 3
Growth protocol Tobacco transgenic lines corresponding to the T4 generation of our previously published DcLCYB1-expressing lines were used in this work (Moreno et al., 2016). Tobacco (Nicotiana tabacum cultivar Xanthi NN) wild type and transgenic lines were raised from seeds germinated in Petri dishes containing MS medium supplemented with 30 g/L sucrose (Murashige and Skoog, 1962) and kanamycin (100 mg/mL).
Extracted molecule total RNA
Extraction protocol Total RNA was extracted from frozen powder of tobacco leaves of 5-week-old T3 plants using the Nucleo Spin RNA extraction kit (Macherey-Nagel, Germany). Genomic DNA traces were eliminated by a 15 min DNase I treatment.
QuantSeq 3’ mRNA-Seq services were performed for the wild type and transgenic DcLCYB1 line L14, L15 and L16, at Lexogen GmbH (Vienna, Austria). 500 ng total RNA were used as input for generating sequencing-ready libraries using the QuantSeq FWD 3’ mRNA-Seq library preparation kit (Lexogen, Vienna, Austria; SKU 015). Libraries were characterized by microcapillary electrophoresis and fluorometry prior to pooling and sequencing.
A sequencing yield of 572,9M-demultiplexed reads was obtained from a single run on a NextSeq 500 device in SR75 HO mode
 
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Illumina NextSeq 500
 
Data processing Integrated data analysis was performed by Lexogen (STAR aligner) where reads were mapped against the Nitab 4.5 reference genome and read-counts were determined.
Differential expression was computed using DESeq2 in R (Love et al., 2014).
Genome_build: Nitab 4.5 reference genome (ftp://ftp.solgenomics.net/genomes/Nicotiana_tabacum/edwards_et_al_2017/)
Supplementary_files_format_and_content: Default STAR output files (ASCII format, tab delimited): Gene identifier, Read count (unstranded RNA-seq), Read count (counts for the 1st read strand aligned with RNA), Read count (counts for the 2nd read strand aligned with RNA)
 
Submission date Sep 05, 2020
Last update date Feb 24, 2021
Contact name Juan Camilo Moreno Beltran
E-mail(s) juankmilo86@gmail.com
Organization name MPI for Molecular Plant Physiology
Department Molecular Physiology
Lab Skirycz
Street address Am Muehlenberg 1
City Potsdam
State/province Brandenburg
ZIP/Postal code 14476
Country Germany
 
Platform ID GPL23495
Series (1)
GSE157541 A multiOMICs approach shed light on the higher yield phenotype and abiotic stress tolerance in tobacco lines expressing the carrot lycopene β-cyclase1
Relations
BioSample SAMN16066989
SRA SRX9081901

Supplementary file Size Download File type/resource
GSM4769700_9RNA15.3.gene_id_count.tab.gz 144.5 Kb (ftp)(http) TAB
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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