title: Detection of miRNA in Cultured Cells or Xenograft Tissues of Breast Cancer.
abstract: MicroRNA (miRNA) analysis has evolved over the past two decades to become a highly specialized field with broad-reaching applications across a multitude of diseases and cellular processes. The choice of an applicable approach for miRNA quantification will depend on a variety of factors such as cost, time constraints, and throughput. Here, we describe the methods of total RNA isolation, AGO2-bound RNA isolation, miRNA polyadenylation, miRNA-cDNA synthesis, and quantitative real-time polymerase chain reaction for the detection of known miRNAs in cultured cells or xenograft tissues of breast cancer.
authors.affiliation: Departments of Pathology and Laboratory Medicine, University of Tennessee Health Science Center, 19 South Manassas Street, Memphis, TN, 38163, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Brown
authors.affiliation: Departments of Pathology and Laboratory Medicine, University of Tennessee Health Science Center, 19 South Manassas Street, Memphis, TN, 38163, USA. mfan2@uthsc.edu.
authors.forename: Meiyun
authors.initials: M
authors.lastname: Fan
mesh_terms: D000818:Animals; D060565:Argonaute Proteins / Q000378:metabolism; D001943:Breast Neoplasms / Q000473:pathology*; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000096:biosynthesis; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D026723:Polyadenylation; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: C466422:AGO2 protein, human; D060565:Argonaute Proteins; D018076:DNA, Complementary; D035683:MicroRNAs
keywords: AGO2 immunoprecipitation; Breast cancer; RNA isolation; miRNA detection; qPCR