| 高效液相色谱法分析逆转录聚合酶链反应产物 |
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| 引用本文： | 廖杰,赵玉兰,董芳霆,杨军,郝秀华. 高效液相色谱法分析逆转录聚合酶链反应产物[J]. 色谱, 1999, 17(5): 491-492 |
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| 作者姓名： | 廖杰 赵玉兰 董芳霆 杨军 郝秀华 |
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| 作者单位： | [1]解放军总医院医学实验测试中心 [2]军事医学科学院国家生物医学分析中心 |
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| 摘 要： | 建立了分离逆转录聚合酶链反应（ＲＴ ＰＣＲ）产物的高效液相色谱方法，反应液直接进样，用ＴＳＫｇｅｌＤＥＡＥ ＮＰＲ柱分离，Ｔｒｉｓ ＨＣｌ缓冲溶液（ｐＨ９０） 氯化钠线性梯度洗脱，于２６０ｎｍ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（ＰＭＮ）磷脂酶Ａ２ｍＲＮＡ的表达。
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| 关 键 词： | 高效液相色谱法 磷脂酶A_2 逆转录聚合酶链反应 |
High Performance Liquid Chromatography for Analysis of Reverse Transcription Polymerase Chain Reaction (RT|PCR) Products |
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| Liao Jie ,Zhao Yulan ,Dong Fangting ,Yang Jun ,Hao Xiuhua. High Performance Liquid Chromatography for Analysis of Reverse Transcription Polymerase Chain Reaction (RT|PCR) Products[J]. Chinese journal of chromatography, 1999, 17(5): 491-492 |
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| Authors: | Liao Jie Zhao Yulan Dong Fangting Yang Jun Hao Xiuhua |
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| Affiliation: | Medical Experiment & Analysis Center of PLA General Hospital, Beijing 100853, China. |
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| 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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