Screening for miRNA Expression Changes Using Quantitative PCR (Q-PCR)

In this section, we describe the use of Applied Biosystems TaqMan Array microRNA Card Set 3.0 to identify miRNA expression in a given RNA sample. This array set includes an array “A” and an array “B” which each have 384 wells that contain specific forward, reverse, and probe oligoinucleotides for measuring the expression of individual miRNAs during a Real-Time PCR (Q-PCR) reaction. Array “A” includes assays for profiling the comparatively higher expressed and better characterized miRNAs. Presently there are 1,048 mature miRNAs annotated in miRBase (release 16). The relatively small amount of miRNAs in comparison to protein-coding genes makes this format a viable option for measuring genome-wide miRNA expression changes. The Applied Biosystems TaqMan miRNA array set 3.0, which includes two separate arrays, can profile 754 miRNAs.