Comparing Methods for Analyzing RNA from Whole Blood
Author Information
Author(s): Kristina Vartanian, Rachel Slottke, Timothy Johnstone, Amanda Casale, Stephen R Planck, Dongseok Choi, Justine R Smith, James T Rosenbaum, Christina A Harrington
Primary Institution: Oregon Health & Science University
Hypothesis
Which RNA preparation and labeling methods yield the most accurate and reproducible microarray results from whole blood samples?
Conclusion
Methods that reduce the impact of globin mRNA transcripts can significantly improve the sensitivity of DNA microarray expression profiling assays for whole blood samples.
Supporting Evidence
- Microarray targets generated with methods that reduce globin mRNA levels detect more transcripts.
- Method-specific differences in correlation with quantitative PCR were observed.
- Freezing blood samples had little impact on RNA expression profiles.
Takeaway
This study looked at different ways to prepare RNA from blood to see which method works best for finding gene information. They found that some methods help get better results by reducing unwanted signals from globin RNA.
Methodology
Four methods for preparing microarray targets from whole blood RNA were compared: no treatment, GLOBINclear treatment, globin PNA treatment, and NuGEN Ovation system.
Potential Biases
Potential bias due to the selection of healthy donors and the specific methods used for RNA extraction and analysis.
Limitations
The study may not generalize to all populations or conditions, and the methods may require specific RNA amounts that are not feasible in some studies.
Participant Demographics
Healthy human donors, specific demographics not detailed.
Digital Object Identifier (DOI)
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