Abstract
MicroRNAs (miRNAs), a newly discovered class of noncoding genes, have diagnostic and prognostic potential for various diseases, most notably for cancer. Microarray-based hybridization has proven to be a powerful technique for miRNA profiling. Formalin-fixed paraffin-embedded (FFPE) tissues are valuable samples for the study of human cancer, since they are generally retrieved with extensively documented clinico-pathological histories. Isolating nucleic acids from archived samples has the potential to unlock a wealth of additional information, facilitating the study of human cancer at the molecular level. In this chapter, we describe one of the potential methods to isolate miRNAs from FFPE.
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Guanidine-thiocyanate in Binding Buffer is an irritant. Always wear gloves and follow standard safety precautions to minimize contact when handling. Do not let these buffers touch your skin, eyes, or mucous membranes. If contact does occur, wash the affected area immediately with large amounts of water; otherwise, the reagent may cause burns. If you spill the reagent, dilute the spill with water before wiping it up. Never store or use the Binding Buffer near human or animal food. Always wear gloves and follow standard safety precautions when handling these buffers. Guanidine-thiocyanate in Binding Buffer can form toxic gases when combined with bleach or acid. If a spilled sample containing this solution is potentially infectious, do not directly add bleach for decontamination.
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Kit requires only one tissue section of 5–10 μm thickness.
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It is possible to modify this protocol in order to use only one column; however, this may lead to higher DNA contamination.
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To minimize the pipetting steps, a 2 ml microcentrifuge tube can be used instead of the collection tube.
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To increase RNA concentration of your sample, elution with 50 μl Elution Buffer is possible.
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© 2011 Springer-Verlag Berlin Heidelberg
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Cirombella, R., Vecchione, A. (2011). MicroRNA Extraction from Formalin-Fixed Paraffin-Embedded Tissues. In: Stanta, G. (eds) Guidelines for Molecular Analysis in Archive Tissues. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-17890-0_15
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DOI: https://doi.org/10.1007/978-3-642-17890-0_15
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