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[High performance liquid chromatography for analysis of reverse transcription polymerase chain reaction (RT-PCR) products].

Jie Liao, Yulong Zhao, Fen Dong, J Yang, Xin Hao

Abstract

The polymerase chain reaction is a powerful method for amplifying specific DNA sequences in vitro. Reverse transcribing mRNA into cDNA expands the use of PCR to monitor mRNA expression in biological system. A method for the analysis of RT-PCR products by HPLC was developed. The separation was performed on a nonporous ion exchange resin column with gradient elution of sodium chloride in 20 mmol/L Tris-HCl buffer (pH 9.0) at a flow rate of 1.0 mL/min and the detection wavelength was 260 nm. lambda-DNA-Hind III digest and a series of RT-PCR products were analyzed for studying the mRNA expression of secreted phospholipase A2 after being injured.

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Available abstract

Key concepts: Chromatography, Reverse transcriptase, Chemistry, Molecular biology, Polymerase chain reaction, Reverse transcription polymerase chain reaction, Complementary DNA, Polymerase