|Table of Contents|

Stem-Loop Real-Time Quantitative PCR for Quantification of miRNA-421 by Specific Primers(PDF)

《南京师大学报(自然科学版)》[ISSN:1001-4616/CN:32-1239/N]

Issue:
2012年02期
Page:
83-88
Research Field:
生命科学
Publishing date:

Info

Title:
Stem-Loop Real-Time Quantitative PCR for Quantification of miRNA-421 by Specific Primers
Author(s):
Zhao LiYang YangWen Chuanjun
Institute of Molecular Cell Biology,School of Life Sciences,Nanjing Normal University,Nanjing 210046,China
Keywords:
stem-loop RT-PCRmiRNA-421primers design
PACS:
Q503
DOI:
-
Abstract:
This study was to establish a method of quantitative detecting miRNA-421 by stem-loop RT-PCR,and to evaluate the feasibility of this method in quantitative assay miRNA. miRBase database was used to get the mature sequence of miRNA-421. 6 primers were designed,including 3 primers for miRNA-421 specific reverse transcription,2 forward and 1 general reverse primers for quantitative PCR,respectively. Different RT primers and forward primers were paired in Q-PCR. RNA after 10 times dilution was reverse transcribed and then detected by Q-PCR. Amplification curves and melting curves were analyzed to identify the primers with high specificity and efficiency of amplification. In order to further validate this primer,miRNA-421 was over-expressed in HepG2 and detected by Q-PCR. 2 primers,421RT7 and 421F19 with high specificity and efficiency of amplification were obtained using stem-loop RT-PCR. Stem-loop RT-PCR provided a method that enabled fast,accurate and sensitive detection of miRNA.

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Last Update: 2013-03-11