Gene expression profiling of primary tumor cell populations using laser capture microdissection, RNA transcript amplification, and GeneChip microarrays.
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.
Luzzi
Veronica I
VI
Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA.
Holtschlag
Victoria
V
Watson
Mark A
MA
D000818:Animals; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D009369:Neoplasms / Q000737:chemistry / Q000235:genetics* / Q000473:pathology*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D018075:RNA, Complementary / Q000032:analysis* / Q000302:isolation & purification; D012334:RNA, Neoplasm / Q000032:analysis* / Q000302:isolation & purification
D018075:RNA, Complementary; D012334:RNA, Neoplasm