title: A melting method for RNA extraction from the mucosal membrane of the mouse middle ear.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors: Department of Otorhinolaryngology, Yonsei University College of Medicine, Seoul, Korea.
authors: Young Joon
authors: YJ
authors: Seo
authors: Department of Otorhinolaryngology, Yonsei University College of Medicine, Seoul, Korea.
authors: Sung Huhn
authors: SH
authors: Kim
authors: Department of Otorhinolaryngology, Yonsei University College of Medicine, Seoul, Korea.
authors: In Seok
authors: IS
authors: Moon
authors: Department of Otorhinolaryngology, Yonsei University College of Medicine, Seoul, Korea. jychoi@yuhs.ac.
authors: Jae Young
authors: JY
authors: Choi
mesh_terms: D000818:Animals; D004432:Ear, Middle / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008855:Microscopy, Electron, Scanning; D055271:Mucin 5AC / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013053:Spectrophotometry
chemical_list: C524679:Muc5ac protein, mouse; D055271:Mucin 5AC; D012333:RNA, Messenger
keywords: Melting; RNA extraction; middle ear mucosa; mouse