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

作者：Patil Kim Vikhe; Ion Barbara Can; Cederroth Christopher R^*

来源：Hearing Research, 2015, 325: 42-48.

DOI：10.1016/j.heares.2015.03.008

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 mu g 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.

• 出版日期2015-7