title: Single-cell laser-capture microdissection and RNA amplification.
abstract: Generating gene-expression profiles from laser-captured cells requires the successful combination of laser-capture microdissection, RNA extraction, RNA amplification, and microarray analysis. To permit single-cell gene-expression profiling, the RNA amplification method has to be sufficiently powerful to bridge the gap between the amount of RNA available from a single cell to what is required by the microarray, a gap that spans 5 to 6 orders of magnitude. This chapter focuses on the amplification of RNA using a two-round T7 RNA amplification method. The protocols described are adapted for laser-captured material and have been used to generate gene expression profiles from single laser-captured cells.
authors.affiliation: Pharmaceutical Research & Development, Johnson & Johnson, San Diego, CA, USA.
authors.forename: Fredrik
authors.initials: F
authors.lastname: Kamme
authors.forename: Jessica
authors.initials: J
authors.lastname: Zhu
authors.forename: Lin
authors.initials: L
authors.lastname: Luo
authors.forename: Jingxue
authors.initials: J
authors.lastname: Yu
authors.forename: Da-Thao
authors.initials: DT
authors.lastname: Tran
authors.forename: Bernhard
authors.initials: B
authors.lastname: Meurers
authors.forename: Anton
authors.initials: A
authors.lastname: Bittner
authors.forename: Karin
authors.initials: K
authors.lastname: Westlund
authors.forename: Susan
authors.initials: S
authors.lastname: Carlton
authors.forename: Jackson
authors.initials: J
authors.lastname: Wan
mesh_terms: D004926:Escherichia coli / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods; D007834:Lasers; D042282:Microdissection / Q000295:instrumentation / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D012313:RNA / Q000235:genetics*
chemical_list: D012313:RNA