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Open Access Research article

Factors affecting the yield of microRNAs from laser microdissectates of formalin-fixed tissue sections

Santosh Kumar Patnaik*, Eric Kannisto and Sai Yendamuri

Author Affiliations

Department of Thoracic Surgery, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY 14203, USA

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BMC Research Notes 2012, 5:40  doi:10.1186/1756-0500-5-40

Published: 19 January 2012

Abstract

Background

Quantification of microRNAs in specific cell populations microdissected from tissues can be used to define their biological roles, and to develop and deploy biomarker assays. In this study, a number of variables were examined for their effect on the yield of microRNAs in samples obtained from formalin-fixed paraffin-embedded tissues by laser microdissection.

Results

MicroRNA yield was improved by using cresyl violet instead of hematoxylin-eosin to stain tissue sections in preparation for microdissection, silicon carbide instead of glass fiber as matrix in RNA-binding columns, and overnight digestion of dissected samples with proteinase K. Storage of slides carrying stained tissue sections at room temperature for up to a week before microdissection, and storage of the microdissectates at room temperature for up to a day before RNA extraction did not adversely affect microRNA yield.

Conclusions

These observations should be of value for the efficient isolation of microRNAs from microdissected formalin-fixed tissues with a flexible workflow.

Keywords:
Cresyl violet; Formalin-fixed tissue; Hematoxylin-eosin; Laser microdissection; MicroRNA; RNA isolation