高效液相色谱法分析逆转录聚合酶链反应产物

High Performance Liquid Chromatography for Analysis of Reverse Transcription Polymerase Chain Reaction (RT\|PCR) Products

建立了分离逆转录聚合酶链反应（ Ｒ Ｔ Ｐ Ｃ Ｒ）产物的高效液相色谱方法，反应液直接进样， 用 Ｔ Ｓ Ｋ ｇｅｌ Ｄ Ｅ Ａ Ｅ Ｎ Ｐ Ｒ 柱分离， Ｔｒｉｓ Ｈ Ｃｌ 缓冲溶液（ｐ Ｈ ９０）氯化钠线性梯度洗脱，于 ２６０ ｎｍ 处检测。用所建立的方法分析了大鼠肠缺血／再灌注损伤后外周血中性粒细胞（ Ｐ Ｍ Ｎ）磷脂酶 Ａ２ ｍ Ｒ Ｎ Ａ 的表达。

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. λ\|DNA\|Hind Ⅲ digest and a series of RT\|PCR products were analyzed for studying the mRNA expression of secreted phospholipase A 2 after being injured.