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Laser microdissection and nucleic acid purification - a Leica - QIAGEN ,,, cooperation

olation and analysis of high-quality DNA and RNA from laser-microdissected samples. The protocols are designed for use with either cryosectioned or formalin-fixed tissues.
Formalin, which is commonly used to fix tissue samples, can cause significant degradation of RNA and DNA in the fixed samples, which affects purification and downstream analysis. Depending on the fixation protocol, the age of the samples, the staining procedure, and the storage conditions used, RNA and DNA may be highly fragmented into pieces smaller than 300 nucleotides, thus limiting the size of fragments isolated. DNA quality from formalinfixed tissues can be significantly improved by cleanup with the QIAquick PCR Purification Kit, as described in the optimized protocol (Figure 2). Flash-frozen, cryosectioned tissues provide higher-quality nucleic acids. For RNA purification and analysis, optimized protocols using the RNeasy Mini Kit are available from QIAGEN. As with DNA isolation, flashfrozen, cryosectioned tissues are preferable
for isolation of RNA. Since the RNeasy procedure removes RNA smaller than 200 nucleotides, this can lead to an overall loss in yield if the RNA is highly degraded. TaqMan analysis demonstrates the better quality of RNA from flash-frozen tissues, as indicated by earlier detection and a lower threshold cycle (Figure 3A and 3B).
Figure 1 Sections of tissue, before (top panel) and after (middle panel) laser microdissection using the Leica AS LMD System, and image of dissected material in the collection tube using inspection mode (bottom panel).
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