Duplex Real-Time PCR Assay for Quantifying Mitochondrial DNA Deletions in Laser Microdissected Single Spiral Ganglion Cells
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.
Key wordsPresbycusis Human temporal bones Mitochondrial DNA deletions Duplex real-time PCR assay Laser microdissection
This work was supported by the Bloom Temporal Bone Laboratory Endowment and a 2009 Deafness Research Foundation Grant provided by the Burch-Safford Foundation to A.M.
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