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Sample GSM8346446 Query DataSets for GSM8346446
Status Public on Jul 01, 2024
Title S47 PLC7_OE shoot drought replicate2
Sample type SRA
 
Source name shoot
Organism Arabidopsis thaliana
Characteristics tissue: shoot
genotype: PLC7_OE
treatment: drought
Treatment protocol 7 days on vermiculite with or without growth media
Growth protocol 9 days on agar, 12 days on vermiculite with growth media
Extracted molecule polyA RNA
Extraction protocol Plant tissues were grinded with mortar and pestle in liquid nitrogen. RNA used for sequencing was extracted using the RNeasy Plant Mini Kit from Qiagen, following the manufacturer’s instructions. The quality of the RNA, i.e. RNA integrity, for sequencing was checked by RIN measurement by ‘Dutch Genomics Service and Support Provider’ (MAD, Science Park, University of Amsterdam, The Netherlands), for which all root samples scored at least 6.5, and all shoot samples at least 9.0.
The total RNA was mixed with ERCC RNA Spike-In Control mix 1 (Thermo Fisher Scientific) and afterwards a poly- A enrichment was performed using Dynabeads mRNA DIRECT Micro Purification Kit (Invitrogen). With the Ion Total RNA-Seq kit v2 and the Ion Xpress RNA-Seq barcoding kit, bar-coded RNA libraries were generated by following the manufacturer’s instructions (Thermo Fisher Scientific)
 
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Ion Torrent Proton
 
Data processing Data was cleaned using FastQC, filtering out all reads smaller than 25 nucleotides and any reads where >75% of the read had a quality score of 17 or less
Reads were aligned to the TAIR10_cdna_20101214_updated reference genome using the Kallisto package
For the number of alignments, 500 was chosen.
The Sleuth package (Pimentel et al., 2017) was used to determine differential gene expression (DEG), the Wald-test was used to determine fold-change difference between conditions or genotypes. Significance was deemed at q<0.05 as determined by the Likelihood-ratio test.
Assembly: TAIR10_cdna_20101214
Supplementary files format and content: abundance.tsv contain est count and tpm for each arabidpsis gene.
Supplementary files format and content: abundance.h5 contain est count and tpm for each of the 500 allignments performed by kallisto. Can be used by the Sleuth package to get average and confidence intervals for the est_counts and tpm
 
Submission date Jun 24, 2024
Last update date Jul 01, 2024
Contact name Max van Hooren
E-mail(s) max.v.hooren@gmail.com
Organization name University of Amsterdam
Department Plant Cell biology
Street address Science park 904
City Amsterdam
State/province Noord Holland
ZIP/Postal code 1098 XH
Country Netherlands
 
Platform ID GPL20696
Series (1)
GSE270544 Transcriptome analyses of Arabidopsis lines overexpressing distinct PLC genes to explore their contribution to drought tolerance
Relations
BioSample SAMN42017484
SRA SRX25034898

Supplementary file Size Download File type/resource
GSM8346446_S47.abundance.h5 63.0 Mb (ftp)(http) H5
GSM8346446_S47.abundance.tsv.gz 520.3 Kb (ftp)(http) TSV
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Raw data are available in SRA

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