High-Throughput Sample Preparation for Gene Expression Profiling and in Vitro Target Validation

The demand for high-throughput RNA isolation has been dramatically increasing with wide applications of RNAi, expression profiling, and molecular diagnosis. A comparison of various RNA isolation methods that have been adapted to high-throughput platforms, focusing on consistently high yield and quality of isolated RNA, reduction of cross-contamination, and simplicity and robustness of the protocol is presented. The streamlining of RNA isolation with RNA quantification by qRT-PCR and amplification for microarray analysis is also discussed. In general, a microspheric bead-based approach results in more consistent RNA recovery than glass fiber filter-based RNA isolation method, and RNA can be eluted in a smaller volume. This is because beads can be fully re-suspended in solution to enable more thorough mixing, washing, and elution, whereas the glass fiber matrix is fixed in a filter plate. (JALA 2004;9:140-5)