description: Protocols: To obtain synchronized populations, gravid adults were treated with hypochlorite and eggs were allowed to hatch in M9 overnight. ~ 700 L1 larvae per strain were plated on 10 cm NG agar plates seeded with concentrated E. coli OP50 and cultured at the required temperatures (GSC(-) and corresponding GSC(+) strains at 25 °C from L1 for 24 h, then shift to 20 °C, to eliminate germ cells [temperature-sensitive glp-1(e2144ts)-mutation]; all other strains 20 °C throughout entire experiment). At the L4-stage, 500 worms per strain were transferred to two E. coli OP50-seeded 6 cm NG agar plates supplemented with 20 µM FUDR to inhibit germ cell proliferation and progeny production. At day 2 of adulthood, worms were harvested by washing them off their plates with M9. After additional washing with M9 and RNAse-free water, worms were suspended in 1 ml Trizol, snap-frozen in liquid nitrogen and stored at -80 °C until RNA-extraction. RNA was extracted using Trizol and cleaned up with the Monarch® Total RNA Miniprep Kit or RNA Cleanup Kit (New England Biolabs) according to the manufacturer's instructions. RNA library preparation was performed by Eurofins Genomics (Konstanz, Germany) using proprietary protocols and NEBNext Ultra II Directional RNA library preparation kit for Illumina.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2021-11-25
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2021-11-25
attributes.tag: External Id
attributes.value: SAMEA9926466
attributes.tag: INSDC center alias
attributes.value: Institute for Biomedical Aging Research, University of Innsbruck
attributes.tag: INSDC center name
attributes.value: Institute for Biomedical Aging Research, University of Innsbruck
attributes.tag: INSDC first public
attributes.value: 2021-11-25T00:18:17Z
attributes.tag: INSDC last update
attributes.value: 2021-11-25T00:18:17Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-10898:gsc(-)_mbk-1(-)_3
attributes.tag: age
attributes.value: 2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: glp-1(e2144ts) III; mbk-1(pk1389) X
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: whole organism
attributes.tag: sample name
attributes.value: E-MTAB-10898:gsc(-)_mbk-1(-)_3
attributes.tag: sex
attributes.value: hermaphrodite
attributes.tag: strain
attributes.value: CF4165