Abstract

Isothermal Parallel Amplification of microRNAs and Microarray-based Detection

Thomas Brandstetter, Senior Scientist, University Freiburg

As part of molecular diagnostics, e.g. for breast cancer therapy, simultaneous amplification of a broad spectrum of microRNA (miRNA) is required. Nevertheless, the number of miRNA molecules that can be analyzed in one assay is limited. This fact is based on the formation of primer dimers during the amplification reaction [1]. Here, an isothermal multi-parametric miRNA amplification method based on universal NASBA is presented [2]. The universal NASBA is a method, which was expanded to boost the multiplex capacity of standard NASBA reactions significantly. This method enables parallel amplification of multiple miRNA sequences. The analysis of the amplicons is performed by using custom-made DNA microarrays [3], which allows to combine the sensitivity and specificity of miRNA amplification and the multiplex-capacity of microarrays.