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缓冲液pH值对肽核酸压电基因传感器阵列杂交效应的影响

Effects of pH values of buffer on hybridization efficiency with PNA piezoelectric gene sensor array

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【作者】 陈鸣府伟灵刘明华张波蒋天伦陈庆海汤万里徐世军蔡国儒

【Author】 CHEN Ming, FU Wei ling, LIU Ming hua, ZHANG Bo, JIANG Tian lun, CHEN Qing hai,TANG Wan li, XU Shi jun, CAI Guo ru (Department of Clinical Laboratories, Southwest Hospital, Third Military Medical University, Chongqing 400038, China)

【机构】 第三军医大学附属西南医院检验科中国嘉陵集团设备工程公司中国电子科技集团公司第二十六研究所第八研究室 重庆400038重庆400038重庆400032重庆400060

【摘要】 目的 探索PBS缓冲液的pH值对肽核酸压电基因传感器阵列杂交效应的影响。方法 压电基因传感器阵列表面先固定上 1 5 μmol L的bis PNA探针 ,然后分别在pH 5 8~ 8 0的PBS缓冲液中和相应的靶序列杂交 ,记录频率的下降值及反应所需的时间。结果 随着pH值从 5 8升到 8 0 ,杂交导致的频率下降值先高后低 ,以pH 6 8为一个分水岭。杂交平衡时间在pH 6 8之前总体上说相差尚不明显 ,但当pH值从 6 8升到 8 0时 ,却骤然增加。结论 bis PNA和靶序列dsDNA的杂交时适用的pH范围很窄 (pH 6 6~ 7 0 )。弱酸环境下 (pH 6 8时 )最为容易 ,pH超过 7 5就不适宜杂交。结果从另一个侧面证实了在bis PNA和dsDNA的杂交过程中“Hoogsteen链第一”的反应机制。

【Abstract】 Objective To investigate the effects of pH values of PBS buffer on hybridization efficiency with peptide nucleic acid (PNA) piezoelectric gene sensor array Methods After 1 5 μmol/L bis PNA probe was immobilized on the surface of gene sensor array, it was hybridized with the corresponding target sequence at pH values of PBS buffer ranging from 5 8 to 8 0 The decrease in frequency and the equilibrium time of reaction were recorded and analyzed Results Frequency change due to hybridization increased first and then decreased when pH values of PBS varied from 5 8 to 8 0 pH 6 8 was the watershed Equilibrium time for hybridization remained almost unchanged when pH value was lower than 6 8, but it increased suddenly when pH value range from 6 8 to 8 0 Conclusion The suitable pH range for smooth hybridization of bis PNA and target dsDNA is quite narrow (about pH 6 6~7 0) but the most suitable one for hybridization is in slightly acid environment, i e pH 6 8 It is not suitable for hybridization if pH value surpasses 7 5 The opinion of "Hoogsteen strand first" during the procedure of bis PNA reacting with dsDNA is confirmed by our results from another point of view

【关键词】 pH值肽核酸基因传感器杂交
【Key words】 pH valuepeptide nucleic acidgene sensorhybridization
【基金】 国家“九五”科技攻关项目 ( 96 A2 3 0 4 0 4 ) ;国家自然科学基金资助面上项目 ( 39870 831) ;国家高技术计划生物技术领域青年基金资助项目 (“86 3”项目 ) ;全军“九五”医学科研计划资助面上项目( 98MO94 ) ;全军“十五”医学科研计划资助面上项目
  • 【文献出处】 第三军医大学学报 ,Acta Academiae Medicinae Militaris Tertiae , 编辑部邮箱 ,2003年15期
  • 【分类号】R446.5
  • 【被引频次】1
  • 【下载频次】75
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