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Sample GSM752575 Query DataSets for GSM752575
Status Public on Oct 13, 2011
Title oan ht M 1
Sample type SRA
Source name Heart
Organism Ornithorhynchus anatinus
Characteristics gender: Male
age: adult
extraction: Standard
rin: 7.3
tissue source: Captured in New South Wales, Australia (provided by Adelaide University)
Extracted molecule polyA RNA
Extraction protocol Total RNA was extracted using the Trizol (Invitrogen) procedure or RNAeasy/RNAeasy Lipid/miRNeasy (Qiagen) column purification kits as indicated. RNA quality was assessed using an Agilent 2100 Bioanalyzer. Sequencing libraries were prepared using the mRNA-Seq Sample Prep Kit (Illumina) according to the manufacturer's instructions. Briefly, polyadenylated RNA was isolated using a poly-dT bead procedure and then chemically fragmented and randomly primed for reverse transcription. After second-strand synthesis, the ends of the double-stranded cDNA were repaired. After 3'-end adenylation of these products, Illumina Paired-End Sequencing adapters were ligated to the blunt ends of the cDNA fragments. Ligated products were run on gels; 250-300 bp fragments were excised and then PCR-amplified (15 cycles). After column-purification, qualities of the resulting libraries were assessed using Agilent 2100 Bioanalyzers. Potential influences on RNA sequencing results due to different experimenters preparing the libraries were ruled out on the basis of RNA- Seq data comparisons of replicate libraries prepared by the different experimenters (i.e., expression levels derived from replicate libraries were highly correlated: rho > 0.99). The RNA-Seq libraries were each sequenced with 76 (101) cycles in at least one lane of the Illumina Genome Analyzer IIx platform according to the manufacturer's specifications. Technical replicates (i.e., sequencing the same library on different machines) were performed to rule out potential biases during the sequencing step (i.e., expression levels between technical replicates were highly correlated: rho > 0.98). After sequencing, we processed the fluorophore intensity files with the Ibis base caller (version 1.11), in addition to applying the standard Illumina base calling algorithms. As illustrated in Supplementary Note Table 1 and Supplementary Note Figure 1 for a small subset of samples, we found that Ibis significantly increased the number of usable reads and drastically reduced the error rate. This improvement was more pronounced for the sequencing runs processed with early versions of the Illumina pipeline, but remained noticeable even for the latest Illumina release (GA pipeline 1.60). All subsequent analyses were performed on the Ibis-called reads.
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Illumina Genome Analyzer IIx
Description sample name: 478
Data processing for processed data, please refer to the original publication
Submission date Jul 01, 2011
Last update date May 15, 2019
Contact name Henrik Kaessmann
Organization name University of Lausanne
Department Center For Integrative Genomics
Lab Kaessmann
Street address GĂ©nopode
City Lausanne
ZIP/Postal code 1015
Country Switzerland
Platform ID GPL13798
Series (1)
GSE30352 The evolution of gene expression levels in mammalian organs
SRA SRX081886
BioSample SAMN00632136

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data not provided for this record

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