Complementary RNA amplification methods enhance microarray identification of transcripts expressed in the C. elegans nervous system

2008

Comparing RNA Amplification Methods for Gene Expression in C. elegans Neurons

Sample size: 5 publication 10 minutes Evidence: moderate

Author Information

Author(s): Joseph D Watson, Shenglong Wang, Stephen E Von Stetina, Clay Spencer W, Shawn Levy, Phillip J Dexheimer, Nurith Kurn, Joe Don Heath, David M Miller III

Primary Institution: Vanderbilt University

Hypothesis

How do different RNA amplification methods affect the identification of transcripts in the C. elegans nervous system?

Conclusion

Both IVT and WT-Pico RNA amplification methods produce valid microarray profiles, but they identify different subsets of neuron-enriched transcripts.

Supporting Evidence

The WT-Pico method identified a higher fraction of present calls compared to IVT.
Both methods detected a significant number of neuron-expressed genes.
WT-Pico amplification was more sensitive, requiring less starting RNA than IVT.
Each method identified unique subsets of neuron-enriched transcripts.
Microarray data showed a significant correlation between the two methods.
WT-Pico amplification resulted in a greater dynamic range of expression.
Transgenic lines confirmed neuronal expression of genes identified by both methods.
Combined data from both methods provided a more comprehensive view of neural gene expression.

Takeaway

Scientists used two methods to make more copies of RNA from tiny samples of C. elegans neurons, and found that each method found different important genes.