| 改进竞争性RT-PCR法在定量PER1基因表达中的应用 |
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| 引用本文： | 刘姝 蔡彦宁 冯秀丽 温玫 左晓虹 陈彪. 改进竞争性RT-PCR法在定量PER1基因表达中的应用[J]. 基础医学与临床, 2005, 25(5): 409-413 |
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| 作者姓名： | 刘姝 蔡彦宁 冯秀丽 温玫 左晓虹 陈彪 |
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| 作者单位： | 首都医科大学宣武医院老年病医学研究所神经生物学室 北京100053(刘姝，蔡彦宁，冯秀丽，温玫，左晓虹)，首都医科大学宣武医院老年病医学研究所神经生物学室 北京100053(陈彪) |
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| 基金项目： | 国家科技攻关计划(2002BA711A07-04)，国家重点基础研究计划(973)(G2000057005)，北京市自然科学基金重点项目(7031002)，北京市科技新星项目(H020821400190)，北京市卫生局重点学科项目 |
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| 摘 要： | 目的研究使用不同样品稀释液及变性高效液相色谱(DHPLC)梯度条件对定量检测PER1基因表达的影响。方法竞争性RT-PCR与DHPLC结合定量检测基因表达。结果样品稀释液中含有载体可极大提高实验的重复性、准确性；不同DHPLC梯度条件对定量结果无明显影响。结论建立定量PER1基因mRNA表达水平的竞争性RT-PCR系统有助于精确检测微量标本中节律基因表达的变化，探寻生物节律与疾病状态间的关系。
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| 关 键 词： | 竞争性RT-PCR 变性高效液相色谱 PER1基因 |
| 文章编号： | 1001-6325(2005)05-0409-05 |
| 修稿时间： | 2004-11-16 |
Improvement of competitive RT-PCR in quantification of PER1 gene expression |
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| LIU Shu,CAI Yan-ning,FENG Xiu-li,WEN Mei,ZUO Xiao-hong,CHEN Biao. Improvement of competitive RT-PCR in quantification of PER1 gene expression[J]. Basic Medical Sciences and Clinics, 2005, 25(5): 409-413 |
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| Authors: | LIU Shu CAI Yan-ning FENG Xiu-li WEN Mei ZUO Xiao-hong CHEN Biao |
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| Affiliation: | LIU Shu,CAI Yan-ning,FENG Xiu-li,WEN Mei,ZUO Xiao-hong,CHEN Biao~* |
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| Abstract: | ObjectiveTo study the impact of different dilution solution and DHPLC washing condition on quantification. MethodsCompetitive RT-PCR system based on DHPLC was used in order to detect PER1 gene expression. ResultsAddition of carriers significantly improved the accuracy and reproducibility of the experiment. Different washing conditions of DHPLC had no obvious influence on the quantification. ConclusionEstablishment of this system would contribute to the study of circadian rhythm in tissues and explore the relationship between circadian rhythm and diseases. |
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| Keywords: | competitive RT-PCR DHPLC PER1 gene |
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