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JoVE Journal Genetics
Retrospective MicroRNA Sequencing
 

Retrospective MicroRNA Sequencing: Complementary DNA Library Preparation Protocol Using Formalin-fixed Paraffin-embedded RNA Specimens

Article DOI: 10.3791/57471-v • 12:24 min • May 5th, 2018
May 5th, 2018 •
1,2,3, 1,2, 3, 4
1Department of Research, Hackensack University Medical Center, 2Department of Medical Sciences, Seton Hall University, 3Department of Epidemiology and Population Health, Albert Einstein College of Medicine, 4Department of Nephrology and Hypertension, Hadassah - Hebrew University Medical Center

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Formalin-fixed paraffin-embedded specimens represent a valuable source of molecular biomarkers of human diseases. Here we present a laboratory-based cDNA library preparation protocol, initially designed with fresh frozen RNA, and optimized for the analysis of archived microRNAs from tissues stored up to 35 years.

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Retrospective MicroRNA Sequencing Complementary DNA Library Preparation Formalin-fixed Paraffin-embedded RNA Specimens Small RNAs Formalin-fixed Paraffin-embedded Tissues RNA Extraction MicroRNA Profiling Ligation Master Mix Adapter Transfer Denaturation Ligation Enzyme Overnight Incubation
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