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Real-time qRT-PCR assay for the detection of miRNAs using bi-directional extension sequences
- Authors
- Kim, Kyung Jin; Kwak, Jiwon; Lee, Jae-Hoon; Lee, Soo Suk
- Issue Date
- 1-Nov-2017
- Publisher
- Academic Press
- Keywords
- microRNA (miRNA); Bi-directional extension (BDE); RT-qPCR; Reverse transcription; Poly(A) tailing assay
- Citation
- Analytical Biochemistry, v.536, pp 32 - 35
- Pages
- 4
- Journal Title
- Analytical Biochemistry
- Volume
- 536
- Start Page
- 32
- End Page
- 35
- ISSN
- 0003-2697
1096-0309
- Abstract
- Highly specific detection of miRNAs was performed using a novel bi-directional extension (BDE) assay. After reverse transcription, the cDNA was hybridized to a uniquely designed specific BDE sequence; this cDNA-BDE hybrid forms the PCR template. The PCR template was amplified in a SYBR Green-based quantitative real-time PCR. The miR-145 in human brain total RNA could be detected quantitatively in the range of seven orders of magnitude with high linearity and reproducibility. This innovative BDE assay has several performance advantages over the poly(A) tailing method that include lower C-T values, clear gel electrophoresis images, and distinct nucleotide peaks in sequencing chromatograms. (C) 2017 Elsevier Inc. All rights reserved.
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Related Researcher
- Lee, Soo Suk
- College of Medical Sciences (Department of Pharmaceutical Engineering)