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

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.