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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.

es in the tissue samples (different areas were isolated from the same section) or by some variances in sample handling. On average the extraction-/precipitation-based method (manufacturer C) showed a higher RNA recovery than the two column-based methods, which is most probably caused by irreversible RNA-binding at the column material. The average yield (manufacturer C) obtained from a 1000 cell area was approximately 700 pg/l when resolving the pellet in 30 l of water. This would result roughly in a theoretical average RNA recovery rate of approx 21 pg per cell. In contrast to this the yield obtained by the two column-based kits varied between approx. 130 and 380 pg/l (1000 cells; 30 l elution volume). Note that the quantitative results from an analysis with the RNA 6000 Pico kit are somewhat dependent on the salt contentof a sample. Only a rough estimation of the RNA concentration of a sample can be obtained.

RNA quality and yield obtained from tissue stained with different methods
In this test, we compared RNA quality and yield when using different standard histochemical staining methods. Therefore, we dissected areas corresponding to 2000 cells and purified the RNA using the kit of manufacturer C (3 replicates for each sample). Serial tissue sections were stained using either Hematoxylin/Eosin (HE), Nuclear Fast Red (NFR), Methylgreen (MG) or Methylene Blue (MB). In general, all four staining procedures were compatible with the RNA Pico Assay, but significant differences in RNA yield and quality were observed. Best results were obtained using the MG stain (average yield: approx. 3100 pg; highly intact RNA; see fig. 3A). HE (figure 3B) and NFR staining (figure 3C) gave very similar results in terms of RNA yield and quality (average yield: approx. 1600 pg; slightly degraded RNA
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