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A melting method for RNA extraction from the mucosal membrane of the mouse middle ear

Authors
 Young Joon Seo  ;  Sung Huhn Kim  ;  In Seok Moon  ;  Jae Young Choi 
Citation
 YONSEI MEDICAL JOURNAL, Vol.56(2) : 497-502, 2015 
Journal Title
YONSEI MEDICAL JOURNAL
ISSN
 0513-5796 
Issue Date
2015
MeSH
Animals ; Ear, Middle/metabolism* ; Ear, Middle/pathology ; Mice ; Microscopy, Electron, Scanning ; Mucin 5AC/genetics ; Mucin 5AC/metabolism* ; RNA, Messenger/genetics* ; RNA, Messenger/metabolism ; Reverse Transcriptase Polymerase Chain Reaction/methods* ; Spectrophotometry
Keywords
Melting ; RNA extraction ; middle ear mucosa ; mouse
Abstract
PURPOSE: 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.
MATERIALS AND 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.
CONCLUSION: 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.
Files in This Item:
T201502002.pdf Download
DOI
10.3349/ymj.2015.56.2.497
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Otorhinolaryngology (이비인후과학교실) > 1. Journal Papers
Yonsei Authors
Moon, In Seok(문인석) ORCID logo https://orcid.org/0000-0002-3951-5074
Choi, Jae Young(최재영) ORCID logo https://orcid.org/0000-0001-9493-3458
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/140415
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