Berezovski Maxim V, Khan Nasrin
University of Ottawa, Ottawa, ON, Canada.
Methods Mol Biol. 2013;1039:245-59. doi: 10.1007/978-1-62703-535-4_20.
MicroRNAs play an important role in gene regulation and disease etiology and are blood-based biomarkers of diseases. Here, we describe a protein-facilitated affinity capillary electrophoresis (ProFACE) method for ultra-sensitive direct miRNA detection as low as 300,000 molecules in 1 mL of blood serum, using single-stranded DNA binding protein (SSB) and double-stranded RNA binding protein (p19) as separation enhancers. This method utilizes either the selective binding of SSB to a fluorescent single-stranded DNA/RNA probe or the binding of p19 to miRNA-RNA probe duplex.
微小RNA在基因调控和疾病病因学中发挥着重要作用，并且是疾病的血液生物标志物。在此，我们描述了一种蛋白质辅助亲和毛细管电泳（ProFACE）方法，用于超灵敏直接检测微小RNA，在1 mL血清中低至300,000个分子，使用单链DNA结合蛋白（SSB）和双链RNA结合蛋白（p19）作为分离增强剂。该方法利用SSB与荧光单链DNA/RNA探针的选择性结合或p19与微小RNA-RNA探针双链体的结合。
