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Sample GSM5730992 Query DataSets for GSM5730992
Status Public on May 23, 2023
Title Nfr Liver Young Het Full M smp53
Sample type SRA
 
Source name Nfr Liver
Organism Nothobranchius furzeri
Characteristics tissue: Liver
age: 6.5 weeks
genotype: APRT heterozygous
feeding condition: Fully-Fed
Sex: Male
Treatment protocol Livers were quickly removed, flash frozen in liquid nitrogen, and then stored in -80 until RNA extraction (up to 3months)
Growth protocol Male or female killifish were either fed ad libidum, or fasted for 24h before liver extraction. All extraction were performed between 9am and 12pm.
Extracted molecule polyA RNA
Extraction protocol Samples were disrupted by bead beating in 300μl of TriZol (Sigma) and a single 3mm metal bead (Eldan, BL6693003000) using TissueLyzer LT (QIAGEN, #85600) with a dedicated adaptor (QIAGEN, #69980). Beating was performed twice at 50Hz for 2 minutes. RNA extraction was performed with Direct-zol RNA Purification Kits (Zymo). RNA concentration and quality were determined by using an Agilent 2100 bioanalyser (Agilent Technologies).
Library preparation was performed using KAPA mRNA HyperPrep Kit (ROCHE-08105936001) according to the recommended protocols. Library concentrations were measured by Qubit (dsDNA HS, Q32854), and quality was measured by Tape Station (HS, 5067-5584). Libraries were sequenced by NextSeq 500 high output kit V2, 75 cycle single-end (Illumina, 20024906) using a NextSeq 500 machine (Illumina) with ~30 million reads per sample.
 
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Illumina NextSeq 500
 
Description Sample 53
Data processing Quality control and adapter trimming of the fastq sequence files were performed with FastQC (v0.11.8) (Andrews, 2017) and the Trim Galore! (v0.6.4) (Krueger, 2015) for Cutadapt (3.4)(Martin, 2011). Options were set to remove Illumina TruSeq adapters and end sequences to retain high-quality bases with phred score >20 and a remaining length >20-bp. Successful processing was verified by re-running FastQC.
Reads was mapped and quantified to the killifish genome Nfu_20140520 (Reichwald et al., 2015; Valenzano et al., 2015) using STAR 2.7.6a (Dobin et al., 2013)
Genome_build: Nfu_20140520
Supplementary_files_format_and_content: Matrix table with raw gene counts for every gene and every sample
 
Submission date Dec 13, 2021
Last update date May 23, 2023
Contact name Itamar Harel
E-mail(s) itamarh@mail.huji.ac.il
Organization name The Hebrew University of Jerusalem
Department Genetics
Lab Harel
Street address Givat Ram
City Jerusalem
ZIP/Postal code 9190401
Country Israel
 
Platform ID GPL26990
Series (1)
GSE190757 Genetic perturbation of AMP biosynthesis extends lifespan and restores metabolic health in a naturally short-lived vertebrate
Relations
BioSample SAMN23970512
SRA SRX13395464

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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