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Accession IconSRP091576

Polysome-seq in zebrafish early embryonic development

Organism Icon Danio rerio
Sample Icon No Downloadable Samples
Technology Badge IconIllumina HiSeq 2500

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We followed the polysomal association of maternal and early zygotic transcriptome over the first few hours of embryonic development, prior to and after MBT. We isolated polysome-associated (bound) and non-polysome-associated (unbound) mRNAs using sucrose gradient centrifugation followed by size fractionation. Using next generation sequencing (RNA-seq), we profiled the transcriptome in polysome-bound and unbound fractions. Our analysis revealed distinct dynamics of polysome association of cytoplasmically polyadenylated maternal mRNAs. Overall design: rRNA-depleted RNA was extracted from pooled embryos of desired stages (Egg, 1-cell, 16-cells, 128-cells, 3.5hpf and 5.3hpf) and subjected to polysome profiling to separate between polysome-free and polysome-associated fragments. Unfractionated, rRNA-depleted samples at corresponding stages were used as background control. One RNA-seq library was generated for each sample.
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