Effects of Starting RNA Quantity on Illumina BeadArrays
Author Information
Author(s): Andy G. Lynch, James Hadfield, Mark J. Dunning, Michelle Osborne, Natalie P. Thorne, Simon Tavaré
Primary Institution: University of Cambridge
Hypothesis
What are the costs and consequences of reducing the starting quantity of RNA for the Illumina BeadArray platform?
Conclusion
The BeadArray platform maintains a low false discovery rate even when small amounts of starting RNA are used, but sensitivity drops significantly.
Supporting Evidence
- The study found that starting RNA quantity affects observed intensities.
- Lower RNA quantities lead to a loss of sensitivity but maintain a low false positive rate.
- Reliable results can be obtained with as little as 10 ng of starting RNA.
- Sample sizes should increase to compensate for the loss of signal-to-noise with low RNA quantities.
Takeaway
Using less starting RNA in experiments can still give reliable results, but you might miss some important details.
Methodology
A nine-level dilution design was used to investigate the effect of changing the quantity of starting mRNA on the performance of Illumina BeadArrays.
Potential Biases
Potential bias due to varying RNA quantities affecting sensitivity and expression levels.
Limitations
The study may not fully account for all variables affecting RNA quantity and microarray performance.
Statistical Information
P-Value
p<0.001
Statistical Significance
p<0.001
Digital Object Identifier (DOI)
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