Development of a robust, low cost stem-loop real-time quantification PCR technique for miRNA expression analysis. - Archive ouverte HAL Access content directly
Journal Articles Molecular Biology Reports Year : 2013

Development of a robust, low cost stem-loop real-time quantification PCR technique for miRNA expression analysis.

(1) , (1) , (2) , (3, 4) , (4) , (5) , (6) , (1) , (1)
1
2
3
4
5
6

Abstract

Development of a rapid and accurate quantification method for the detection of microRNAs (miRNAs) has been desired, in particular, when they are differently expressed in normal and pathological conditions. However, various methods for the quantification of small non-coding RNAs as well as miRNAs have been described. These methods mainly include hybridization-based approaches such as primer extension, northern blotting, microarray profiling, and reverse transcription (RT) PCR. Here, we developed a simple and rapid method based on stem-loop primer-based real-time PCR assay for sensitive and accurate detection of mature miRNAs. Initially, a miRNA-specific stem-loop RT primer is used for RT, which is followed by TaqMan real-time PCR assay using specific forward primer in combination with universal reverse primer and TaqMan probe. The assay has shown high sensitivity (≤50 copies/reaction) for miRNA detection in two breast cancer cell lines, MCF-7 and MDA-MB-231. This assay might be implicated as a rapid and cost effective method for the detection of small non-coding RNAs.
Embargoed file
Embargoed file
Ne sera jamais visible
Loading...

Dates and versions

pasteur-00821223 , version 1 (08-05-2013)

Identifiers

Cite

Samira Mohammadi-Yeganeh, Mahdi Paryan, Siamak Mirab Samiee, Masoud Soleimani, Ehsan Arefian, et al.. Development of a robust, low cost stem-loop real-time quantification PCR technique for miRNA expression analysis.. Molecular Biology Reports, 2013, 40 (5), pp.3665-74. ⟨10.1007/s11033-012-2442-x⟩. ⟨pasteur-00821223⟩

Collections

RIIP RIIP_IRAN
95 View
1 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More