High quality RNA extraction of the mammalian cochlea for qRT-PCR and transcriptome analyses.
Hear Res
; 325: 42-8, 2015 Jul.
Article
em En
| MEDLINE
| ID: mdl-25818515
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
RNA
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Cóclea
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Análise de Sequência com Séries de Oligonucleotídeos
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article