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Differential Expression Analysis

Important notes to the user of this service

  • For reliable transcript expressions, a sufficiently high coverage (at least 50 million reads) and a considerable amount of samples (>5) for each class is recommended.
  • If your data is not yet aligned to a reference genome and available in .bam format, our Read-Mapping service will provide adequate input data.

Required input data

  • sequence reads aligned against a reference genome (e.g. hg19 for homo sapiens) in .bam format.
  • assignment of samples to 2 or more named classes (e.g. in a table)

Generated results

  • list of genes expressed differentially between given classes
  • list of transcripts expressed differentially between given classes
  • various summarizing plots (histogram/boxplot of expression distribution in each class, bi-clustered expression heatmap of differentially expressed genes/transcripts against the given classes)

Additional options on request

  • bias correction
  • upper-quartile normalization to improve robustness for input data with low coverage
  • transcript assembly (planned)

Methods & Implementation

Estimation of gene/transcript expression, normalization and detection of differentially expressed genes/transcripts is conducted with cufflinks 1.3.0. Genes and transcripts with a (multiple testing corrected) p-value at most 0.05 are selected as differentially expressed. Summarizing plots are generated with the R package cummeRbund 1.1.3.
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