A Rapid and Real-time Nucleic Acid Detection System by Fluorescence Resonance Energy Transfer and Isothermal RNA Amplification
Date Issued
2006
Date
2006
Author(s)
Chen, Jiann-Mou
DOI
zh-TW
Abstract
In this thesis, we develop a detecting method with high accuracy and high sensitivity for specific RNA sequences in isothermal environment. We construct a low-cost and real-time system for detection specific RNA sequences by using this method.
Isothermal RNA amplification is a powerful tool to amplify specific sequences of RNA. It can successfully amplify as low as 1 ng of total RNA in 90 minutes. Therefore, we integrate isothermal RNA amplification and Fluorescence Resonance Energy Transfer (FRET) methods to real-time detect fluorescent signals for specific RNA sequences. For increase specificity, two hybridization probes which can bind to the same gene are used in our experiments to increase the accuracy. Base on this study, a novel method with higher accuracy and sensitivity for detecting specific RNA sequence in isothermal environment can be achieved.
The results shown that the FRET signal of TRIM28 and UCHL1 genes have been detected in 40-60 min. Therefore, we have successfully setup a new low-cost, real-time detection system, based on combined Isothermal RNA Amplification and FRET.
Subjects
螢光共振能量轉換
恆溫ＲＮＡ擴增
核酸偵測
Fluorescence Resonance Energy Transfer
Isothermal RNA Amplification
Nucleic Acid Detection
Type
thesis
File(s)![Thumbnail Image]()
Loading...
Name
ntu-95-R93921128-1.pdf
Size
23.31 KB
Format
Adobe PDF
Checksum
(MD5):15b98b1024e6ad7b2d60dfe796662add