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Quality assurance of RNA derived from,,,laser microdissected tissue samples,,,obtained by the PALM MicroBeam System,,,using the RNA 6000 Pico LabChip kit.


Gene expression profiling of material isolated by microdissection has become a very popular method for analyzing cellular behavior in a micro scale and is used in a wide range of research and clinical applications. Laser-assisted microdissection allows isolation of RNA from specific cell populations out of their surrounding tissue environment. Usually, the sample yield that can be obtained by this method is too low for quality control of RNA using a standard technique. On the other hand, extraction of high quality RNA is crucial for all subsequent steps and the success of the overall experiment. Since the introduction of the Agilent RNA 6000 Pico LabChip kit, it is possible for the first time to analyze total RNA samples in concentrations down to 200 pg/l, which is in the range of the sample concentration one can expect from microdissected tissue or cells. In this study we analyzed RNA samples that were obtained by laser microdissection and pressure catapulting (LMPC; PALM MicroBeam System) from tissue sections. The influence of different staining procedures and RNA extraction kits on RNA quality was investigated. RNA was tested for quality and approximate yield, as well as the reproducibility within homologous sample replicates. The data revealed clear differences between the different isolation kits and also between the staining procedures that were used.
The analysis of gene expression has become one of the major scientific tools for understanding the behavior of cells in vivo and in vitro and is used in many biological and medical applications. Using microarray hybridization, the abundance of a large number of transcripts can be analyzed in a single experiment. Comparison of different expression patterns enables scientists to correlate the changes in the
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