TY - JOUR
T1 - A melting method for RNA extraction from the mucosal membrane of the mouse middle ear
AU - Seo, Young Joon
AU - Kim, Sung Huhn
AU - Moon, In Seok
AU - Choi, Jae Young
N1 - Publisher Copyright:
© Yonsei University College of Medicine 2015.
Copyright:
Copyright 2015 Elsevier B.V., All rights reserved.
PY - 2015
Y1 - 2015
N2 - 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.
AB - 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.
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U2 - 10.3349/ymj.2015.56.2.497
DO - 10.3349/ymj.2015.56.2.497
M3 - Article
C2 - 25684001
AN - SCOPUS:84922787079
VL - 56
SP - 497
EP - 502
JO - Yonsei Medical Journal
JF - Yonsei Medical Journal
SN - 0513-5796
IS - 2
ER -