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Gene expression profiling of primary tumor cell populations using laser capture microdissection, RNA transcript amplification, and GeneChip microarrays.

Methods in Molecular Biology (Clifton, N.j.)

Animals, Gene Expression Profiling, methods, Humans, Lasers, Microdissection, Neoplasms, chemistry, genetics, pathology, Oligonucleotide Array Sequence Analysis, RNA, Complementary, analysis, isolation & purification, RNA, Neoplasm

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      Abstract

      Gene expression profiling from microdissected cell populations is a powerful approach to explore molecular processes involved in development and solid tumor biology. In this chapter, we detail robust and validated methods for tissue preparation and isolation of high-quality RNA from microdissected cell populations. A protocol is also provided for linear transcript amplification using as little as 10 ng of total RNA to produce labeled cRNA targets for hybridization to GeneChip high-density oligonucleotide microarrays. Particular emphasis is placed on troubleshooting each technical step in the protocol and measures of quality assurance for both RNA isolation and resulting microarray data.

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