By Nobuyoshi Kosaka
microRNAs (miRNAs) are small non-coding RNAs that control quite a few organic phenomena, equivalent to improvement and homeostasis. The dysregulation of miRNA ends up in affliction development, relatively of melanoma. In Circulating MicroRNAs: equipment and Protocols, professional researchers within the box element fresh advances within the isolation, purification and research of circulating miRNAs from numerous resources for study. The publication is split into 3 major subject matters. the 1st part contains the examine of secretory miRNAs in cell-cell verbal exchange, and the second one, the examine of circulating miRNAs in physique fluids. The final describes the radical strategies used to review circulating miRNAs. Written within the hugely profitable Methods in Molecular Biology™ sequence structure, chapters comprise introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, conveniently reproducible laboratory protocols, and key tips about troubleshooting and fending off recognized pitfalls.
Authoritative and practical, Circulating MicroRNAs: tools and Protocols seeks to assist scientists in facing the new advances of RNAi know-how from the bench to the bedside.
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Additional info for Circulating MicroRNAs: Methods and Protocols
1038/ nrc3166 11. Kogure T, Lin WL, Yan IK et al (2011) Intercellular nanovesicle-mediated microRNA transfer: a mechanism of environmental modulation of hepatocellular cancer cell growth. Hepatology 54:1237–1248. 24504 12. Schwarzenbach H, Hoon DS, Pantel K (2011) Cell-free nucleic acids as biomarkers in cancer patients. 1038/nrc3066 13. Almqvist N, Lonnqvist A, Hultkrantz S et al (2008) Serum-derived exosomes from antigen-fed mice prevent allergic sensitization in a model of allergic asthma. x 14.
12. S. , Del Mar, CA, USA), or similar model. 13. Magnetic stir plate. 14. Peristaltic pump. 15. NanoDrop 2000c spectrophotometer (Thermo Scientific). 16. 0 software (NanoSight). 17. Refractometer. 18. RNase A. 19. TRIzol® Reagent (Invitrogen). 20. 8 %, for ACS analysis, stabilized with ethanol. 21. miRNAeasy Mini Kit (Qiagen). 22. Qiacube (Qiagen), optional, for fully automated sample processing using Qiagen spin columns. 23. Agilent 2100 Bioanalyzer (Agilent Technologies Inc). 24. ). 25. Taqman MicroRNA Assays for each specific miRNA (Applied Biosystems).
30. Neon™ Transfection System 10 μl Kit (Invitrogen). 31. Dual-Light® System, chemiluminescent reporter gene assay system for the combined detection of firefly luciferase and β-galactosidase (Applied Biosystems). 32. Assay plates for luminometry, 96-well flat bottom, no lid, white polystyrene. 33. Luminometer. 24 3 Angela Montecalvo et al. Methods Exosomes are purified from culture supernatants of mouse bone marrow (BM)-derived DCs (BMDCs) generated in vitro from BM precursors cultured in complete medium supplemented with GM-CSF and IL-4.