title: Duplex real-time PCR assay for quantifying mitochondrial DNA deletions in laser microdissected single spiral ganglion cells.
abstract: Laser microdissection (LMD) has been used to isolate groups of cells and single cells from numerous tissues. In this chapter, we describe a technique for isolating individual spiral ganglion cells from archival formalin-fixed, celloidin-embedded (FFCE) human temporal bone sections. The DNA isolated from these single cells is suitable for analysis with a duplex real-time polymerase chain reaction (PCR) methodology to quantify the mitochondrial DNA (mtDNA) deletion level present.
authors.affiliation: Section of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Chicago, Chicago, IL, USA. amarkaryan@hei.org
authors.forename: Adam
authors.initials: A
authors.lastname: Markaryan
authors.forename: Erik G
authors.initials: EG
authors.lastname: Nelson
authors.forename: Raul
authors.initials: R
authors.lastname: Hinojosa
mesh_terms: D002469:Cell Separation / Q000379:methods*; D004272:DNA, Mitochondrial / Q000235:genetics* / Q000302:isolation & purification; D003627:Data Interpretation, Statistical; D017353:Gene Deletion; D006801:Humans; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D011304:Presbycusis / Q000235:genetics / Q000473:pathology; D012015:Reference Standards; D013136:Spiral Ganglion / Q000378:metabolism / Q000473:pathology*
chemical_list: D004272:DNA, Mitochondrial