Poly(A)-Tailed Universal Reverse Transcription

Abstract

Zheng’ laboratory from the Beijing Institute of Radiation Medicine (Beijing, People’s Republic of China) and Yan’s laboratory from the Department of Physiology and Cell Biology, the University of Nevada School of Medicine (Reno, NV, USA) independently established the Poly(A)-Tailed Universal Reverse Transcription technique, a simple method to detect the expression of mature miRNAs all in the same year (Mol Biotechnol 32:197–204, 2006; Biochem Biophys Res Commun 351:756–763, 2006). With this method, total RNA is first polyadenylated by poly(A) polymerase, and then cDNA is synthesized by a specific reverse transcriptase (RT) primer and RT, using the poly(A)-tailed total RNA as templates. Next, the cDNA is amplified using a miRNA-specific primer and a universal primer. The expression of several mature miRNAs was assayed by this method. Compared with other RT-PCR methods for miRNA detection, the poly (A)-tailed RT-PCR is a relatively simple, convenient, and accurate method.