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Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and terfa-/- zebrafish embryos

ABSTRACT: Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and terfa-/- zebrafish embryos

PROVIDER: PRJNA726517 | ENA |

REPOSITORIES: ENA

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Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare the early stage of pEGFP-N1 injected zebrafish embros transcriptome profiling (RNA-seq) to microarray and quantitative reverse transcription polymerase chain reaction (qRT–PCR) methods and to evaluate protocols for optimal high-throughput data analysis Methods: we injected the plasmid pEGFP-N1 into zebrafish zygotes by microinjection, and then when the embryos to develop to 1 hour-post-fertilization (hpf), 6 hpf and 12 hpf, these groups was collected respectively by deep sequencing, in triplicate, using Illumina GAIIx. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Results: Bioinformatics analysis of the transcriptome sequencing found that the expression levels of genes related to the apoptosis process (Fig.1A), endogenous immune response and tumor necrosis factor-mediated signaling pathways were abnormal up-regulation, mainly including isg15, foxo3b, phlda3, cdkn1a, zgc:136826, and si:dkey-204l11.1, compared with the non-injected group at 6hpf. These genes are found in somatic cells to deal with foreign plasmids invasion.

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Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and terfa-/- zebrafish embryos

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Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and smyd4 mutant zebrafish cardiac transcriptomes

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