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SRX15045259: GSM6076958: Xenopus laevis Embryos, EU-RNA, 9 hpf, Rep 1 [Bead_9hpf_rep1]; Xenopus laevis; RNA-Seq
4 ILLUMINA (NextSeq 500) runs: 15M spots, 1.2G bases, 475.3Mb downloads

External Id: GSM6076958_r1
Submitted by: Good Lab, Cell and Developmental Biology, University of Pennsylvania
Study: Total and Nascent Transcriptome of Xenopus laevis Embryos at 5-9 hpf (stage 7-9)
show Abstracthide Abstract
To characterize the nascent transcriptome during zygotic genome activation (ZGA) of Xenopus laevis embryos, we microinjected 5-ethynyl uridine (EU) into 1-cell stage embryos and isolated total RNAs from whole embryos at 5, 6, 7, 8 and 9 hours post-fertilization (hpf) at room temperature, respectively, covering the stages of pre-ZGA to widespread ZGA (stage 7-9). To purify nascent transcripts, total RNAs were biotinylated using disulfide biotin azide via click reaction and biotinylated nascent transcripts were purified using streptavidin beads. Libraries were constructed from using the total RNA ('All'), nascent transcripts ('Bead') and the flowthrough after purification of nascent transcripts ('FL'), respectively. To categorize maternal-zygotic (MZ) genes and zygotic-only (Z) genes, total RNAs from eggs were isolated for constructing libraries. All libraries were sequenced on illumina NextSeq 500. Overall design: Two experiments were conducted. In the first experiment, two clutches of embryos at 5-9 hpf (two biological replicates) were used for sequecing 'All' (total RNA), 'Bead' (nascent transcripts) and FL (flowthrough RNAs), repsecitvely. In the second experiment, two clutches of embryos at 5-9 hpf (two biological replicates) were used for sequecing 'Bead' (nascent transcripts) and FL (flowthrough RNAs), repsecitvely, two clutches of unfertilized eggs (teo biilogical replicates) were used for sequencing 'All' (total RNA), and two clutches of embryos at 5 hpf (two biological replicates) were used for sequecing 'All' (total RNA).
Sample: Xenopus laevis Embryos, EU-RNA, 9 hpf, Rep 1 [Bead_9hpf_rep1]
SAMN27962914 • SRS12791414 • All experiments • All runs
Organism: Xenopus laevis
Library:
Name: GSM6076958
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Total RNAs were isolated using the RNeasy Mini Kit (Qiagen), following the instructions provided by the manufacturer. Nascent EU-RNAs were biotinylated using bisulfide biotin azide via click reaction and purified using streptavidin beads following the instructions provided by the Click-iTTM Nascent RNA Capture Kit (Thermo Fisher Scientific, Cat# C10365). The cDNA libraries for 'All' (Total RNA), 'Bead' (Nascent transcripts), 'FL' (Flowthrough RNA) and 'Egg' (Maternal RNA) were prepared using the Universal RNA-seq with NuQuant® kit (NuGEN, Cat# 0364), following the manual provided by the manufacturer. Ribosomal RNAs were depleted using the custom designed AnyDeplete Probe Mix for Xenopus laevis provided by the kit.
Runs: 4 runs, 15M spots, 1.2G bases, 475.3Mb
Run# of Spots# of BasesSizePublished
SRR189691563,857,442320.4M122.4Mb2022-08-23
SRR189691573,613,680300.1M113.9Mb2022-08-23
SRR189691583,868,131321.3M122.4Mb2022-08-23
SRR189691593,700,940307.4M116.6Mb2022-08-23

ID:
21519779

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