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

BMC Research Notes

Sai Yendamuri¹  Eric Kannisto¹  Santosh Kumar Patnaik¹ 

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

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

Others  :  1166764
DOI  :  10.1186/1756-0500-5-40

 received in 2011-10-12, accepted in 2012-01-19,  发布年份 2012

【 摘 要 】

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.

【 授权许可】

   
2012 Patnaik et al; licensee BioMed Central Ltd.

【 图 表 】

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