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Systematic analysis of T7 RNA polymerase based in vitro linear RNA amplification for use in microarray experiments
Jörg Schneider1, Andreas Buness, Wolfgang Huber
1Department of Molecular Genome Analysis, German Cancer Research Centre, Heidelberg 69120, Germany. joerg.schneider@dkfz.de
BMC Genomics
|May 4, 2004
Summary
RNA amplification using the T7 polymerase method is reliable for microarray experiments. This technique overcomes the limited RNA yield from small biopsies without introducing significant systematic bias.
Area of Science:
- Molecular Biology
- Genomics
Background:
- Microarray experiments require substantial high-quality RNA (10-100 microg), often unattainable from small biopsies.
- Existing in vitro RNA amplification methods for microarrays lack systematic bias analysis.
Purpose of the Study:
- To evaluate the T7 RNA polymerase-based RNA amplification method for potential biases in microarray studies.
- To assess the suitability of RNA amplification for routine laboratory use in genomics.
Main Methods:
- Hybridization studies on microarrays were conducted to examine error sources.
- Statistical analysis was performed on parameters relevant to laboratory implementation.
Main Results:
- RNA amplification demonstrated no systematic influence on microarray experiment outcomes.
- While minor variations exist, RNA amplification is reproducible and does not introduce significant systematic bias.
Conclusions:
- The T7 RNA polymerase-based amplification method is a valuable tool for microarray experiments.
- Consistent sample treatment is crucial for reliable results when using RNA amplification.