title: Laser-capture microdissection of renal tubule cells and linear amplification of RNA for microarray profiling and real-time PCR.
abstract: Laser-capture microdissection and transcriptional profiling have enabled compartment- and cell-specific analysis of gene expression in chronic kidney disease, thus facilitating the investigation of pathophysiological associations between glomerular, tubular, and interstitial structures. Due to the pico- and nanogram amounts of RNA isolated from LCM-captured material linear RNA amplification protocols are necessary prior to real-time PCR and microarray analysis. In this chapter, we describe the isolation of renal tubule cells from cryocut sections from routine kidney biopsies, and the isolation and linear amplification of RNA for downstream purposes.
authors.affiliation: Department of Internal Medicine IV (Nephrology and Hypertension), Functional Genomics Research Group, Center of Internal Medicine, Medical University Innsbruck, Innsbruck, Austria.
authors.forename: Susie-Jane
authors.initials: SJ
authors.lastname: Noppert
authors.forename: Susanne
authors.initials: S
authors.lastname: Eder
authors.forename: Michael
authors.initials: M
authors.lastname: Rudnicki
mesh_terms: D018076:DNA, Complementary / Q000737:chemistry; D020869:Gene Expression Profiling / Q000379:methods*; D007684:Kidney Tubules / Q000166:cytology*; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger