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Hear Res. 2015 Jul;325:42-8. doi: 10.1016/j.heares.2015.03.008. Epub 2015 Mar 27.

High quality RNA extraction of the mammalian cochlea for qRT-PCR and transcriptome analyses.

Author information

1
Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, 17177, Sweden. Electronic address: kim.Vikhe.patil@stud.ki.se.
2
Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, 17177, Sweden. Electronic address: Barbara.Canlon@ki.se.
3
Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, 17177, Sweden. Electronic address: christopher.cederroth@ki.se.

Abstract

Molecular investigations of the hearing organ, the cochlea, have been hampered due to the difficulty of isolating pure RNA and in quantities sufficient enough for quantitative real-time RT-PCR or microarray analysis. The complex architecture of the cochlea, the presence of liquids, bone and cartilage tissue, are a major hurdle in obtaining contamination-free RNA to a level that does not affect downstream applications. Here, we present a protocol to extract RNA from the mouse cochlea, with yields and quality suitable for real-time RT-PCR or Affymetrix labeling. In contrast to current methods, such as TRIZOL or column-based extraction, this protocol combines the two and, within 4 h, yields a 2 μg of total RNA from a single pair of adult mouse cochleae. This protocol allows the isolation of RNA molecules from the mammalian cochlea providing access to whole-transcript expression analyses.

PMID:
25818515
DOI:
10.1016/j.heares.2015.03.008
[Indexed for MEDLINE]

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