Journal
BIOTECHNOLOGY LETTERS
Volume 34, Issue 4, Pages 627-633Publisher
SPRINGER
DOI: 10.1007/s10529-011-0813-3
Keywords
microRNA; PolyA RT-PCR; SYBR Green; TaqMan probe
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Funding
- National Key Basic Research Program of China (973 Program) [2011CB504502, 2009CB941704]
- Science and Technology Commission of the Shanghai Municipality Foundation [09DJ1400601]
- Key Projects in the National Science and Technology Pillar Program in the Eleventh Five-year Plan Period, China [2006BAI05A05]
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Quantification of microRNAs (miRNAs) in tissues under normal and pathological conditions is important for elucidating miRNA functions. Based on a PolyA RT-PCR method we have described (J Zhang et al. Biochem Biophys Res Commun 2008 377:136-140), a modified miRNA quantification method was developed and validated using a universal TaqMan probe complementary to the reverse transcript primer. This method effectively detects miRNA expression in cell lines and tissues. The TaqMan probe is more accurate and reliable than the SYBR Green method since it was free from primer dimers. A series of miRNAs were tested in five different mouse tissues: the method differentiated different miRNAs of the same family. This universal TaqMan probe-based PolyA RT-PCR method showed its advantages in precision, simplicity and high-throughput capability compared with other miRNA-detecting methods.
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