Seo Young Joon, Kim Sung Huhn, Moon In Seok, Choi Jae Young
Department of Otorhinolaryngology, Yonsei University College of Medicine, Seoul, Korea.
Yonsei Med J. 2015 Mar;56(2):497-502. doi: 10.3349/ymj.2015.56.2.497.
There is much confusion surrounding the methods of RNA extraction from the middle ear mucosa of mice. In this study, we worked to develop a "melting method," which is faster, purer, and more reliable than other methods in common use.
Thirty-two ears were used for this study. Light microscopy with hematoxylin-eosin staining of the bullae, scanning electron microscopy (SEM), spectrophotometer analysis, and reverse transcription polymerase chain reaction were performed before and after melting the half lateral bullae, which were detached from the temporal bone by using a lateral retroauricular approach.
Each resected half bulla contained a well distributed mucosal membrane. After a TRIzol melting duration of 10-30 minutes, only mucosal marker (MUC5AC) was expressed without bony marker (total osteocalcin). The same results were determined from SEM.
This melting method, compared with stripping and irrigation methods, is effective and offers an easier, more robust approach to extracting RNA from the middle ear mucosal membranes of mice.
从小鼠中耳黏膜提取RNA的方法存在诸多混淆之处。在本研究中，我们致力于开发一种“融化法”，该方法比其他常用方法更快、更纯且更可靠。
本研究使用了32只耳朵。通过耳后外侧入路从颞骨分离出半侧大泡后，在融化前后对大泡进行苏木精-伊红染色的光学显微镜检查、扫描电子显微镜（SEM）检查、分光光度计分析以及逆转录聚合酶链反应。
每个切除的半侧大泡都含有分布良好的黏膜膜。在TRIzol融化持续10 - 30分钟后，仅表达黏膜标志物（MUC5AC），而无骨标志物（总骨钙素）表达。SEM检查也得出了相同结果。
与剥离法和冲洗法相比，这种融化法有效，且为从小鼠中耳黏膜提取RNA提供了一种更简便、更可靠的方法。
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