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Analysis of gene expression profiles on 38 days-old sea bass heads

ABSTRACT: A sea bass oligo microarray platform was used to profile gene expression in whole heads of 38 days-old sea bass affected by prognathism, a skeletal malformation that strongly affects sea bass production. Two different conditions: i) prognathous individuals, and ii) normal individuals were analyzed. For each condition, total RNA was extracted from three (3) independent biological replicates, each consisting of pools of five (5) heads. Statistical analysis with SAM (Significance Analysis of Microarray) didn’t identify any difference in expression patterns between the two groups. Samples were then employed as biological replicates to determine array-to-array reproducibility, the degree of mutual agreement among replicates was estimated using Pearson correlation coefficients on the entire set of expression values. For all pairs of experiments correlation coefficients were always significant (p-value <0.01) and never less than 0.99.

ORGANISM(S): Dicentrarchus labrax

PROVIDER: GSE19041 | GEO | 2010/10/22

SECONDARY ACCESSION(S): PRJNA120529

REPOSITORIES: GEO

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Analysis of gene expression profiles on 38 days-old sea bass heads

Project description:A sea bass oligo microarray platform was used to profile gene expression in whole heads of 38 days-old sea bass affected by prognathism, a skeletal malformation that strongly affects sea bass production. Two different conditions: i) prognathous individuals, and ii) normal individuals were analyzed. For each condition, total RNA was extracted from three (3) independent biological replicates, each consisting of pools of five (5) heads. Statistical analysis with SAM (Significance Analysis of Microarray) didnât identify any difference in expression patterns between the two groups. Samples were then employed as biological replicates to determine array-to-array reproducibility, the degree of mutual agreement among replicates was estimated using Pearson correlation coefficients on the entire set of expression values. For all pairs of experiments correlation coefficients were always significant (p-value <0.01) and never less than 0.99. In this study, we analyzed six (6) samples, three (3) pools of heads dissected from normal sea bass and three (3) dissected from individuals affected by prognathism. Gene expression profiling was performed using the Agilent-019810 Dicentrarchus labrax Oligo Microarray platform (6 arrays, no replicate) based on single-colour detection (Cyanine-3 only). Microarrays were scanned with Agilent scanner G2565BA (barcode on the left, DNA on the back surface, scanned through the glass) at a resolution of 5 microns; all slides were scanned twice at two different sensitivity settings (XDRHi 100% and XDRLo 10%); the scanner software created a unique ID for each pair of XDR scans and saved it to both scan image files. Feature Extraction (FE) 9.5 used XDR ID to link the pairs of scans together automatically when extracting data. The signal left after all the FE processing steps have been completed is ProcessedSignal that contains the Multiplicatively Detrended, Background-Subtracted Signal.

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Comparison of gene expression profile between 58 days-old prognatous sea bass jaws and control ones

Project description:A sea bass oligo microarray platform was used to profile gene expression in mandibles of 58 days-old sea bass affected by prognathism, a skeletal malformation that strongly affects sea bass production. Two different conditions: i) protruding jaws, and ii) normal jaws were used for gene expression analysis. For each condition, total RNA was extracted from four (4) independent biological replicates, each consisting of pools of five (5) jaws. Statistical analysis with SAM (Significance Analysis of Microarray) identified 333 probes (corresponding to 242 unique transcripts) significantly down-regulated in deformed individuals compared to normal ones. In this study, we analyzed eight(8) samples, four (4) pools of jaws dissected from normal sea bass and four (4) pools of jaws dissected from individuals affected by prognathism. Gene expression profiling was performed using the Agilent-019810 Dicentrarchus labrax Oligo Microarray platform (8 arrays, no replicate) based on single-colour detection (Cyanine-3 only). Microarrays were scanned with Agilent scanner G2565BA (barcode on the left, DNA on the back surface, scanned through the glass) at a resolution of 5 microns; all slides were scanned twice at two different sensitivity settings (XDRHi 100% and XDRLo 10%); the scanner software created a unique ID for each pair of XDR scans and saved it to both scan image files. Feature Extraction (FE) 9.5 used XDR ID to link the pairs of scans together automatically when extracting data. The signal left after all the FE processing steps have been completed is ProcessedSignal that contains the Multiplicatively Detrended, Background-Subtracted Signal.

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Project description:Purpose: We used transcriptome sequencing to study the regulated mechanism of VvSUC11,VvSUC12,or VvSUC27 on tomato fruit growth. Methods: the 35S:: VvSUC11, VvSUC12, and VvSUC27 constructs were introduced into transgenic tomatoes (Micro-Tom) by the Agrobacterium-mediated transformation, 5 DPA (days after anthesis) fruit was collected, 3 replicates per group. Extract total RNA, detect RNA quality, library preparation, sequencing, and analyze data. Results:Reads were obtained for each sample using Illumina Hiseq X Ten sequencing. After discarding the low-quality reads, corresponding to over 40 million clean reads obtained from sequencing were mapped on the reference genome of Solanum lycopersicum. High Pearson's correlation coefficients of FPKM distribution between the three biological replicates for each sample were detected (1> R2 > 0.99, P < 0.001) , reflecting the robustness of our library preparation from tomato RNA samples. Compared with WT lines, there were 2459 DEGs in the VvSUC11-overexpressing lines, of which 1283 genes were upregulated and 1176 genes were downregulated. Whereas, there were 1369 DEGs in VvSUC12-overexpressing lines, including 544 upregulated and 825 downregulated genes. In the meanwhile,there were 1777 DEGs in VvSUC27-overexpressing lines, including 623 upregulated and 1154 downregulated genes.

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