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PCR温度特性实验研究与分析
  • ISSN号:1671-7929
  • 期刊名称:《生命科学仪器》
  • 时间:0
  • 分类:Q7[生物学—分子生物学] R4[医药卫生—临床医学]
  • 作者机构:[1]上海理工大学,上海200093, [2]淮海工学院,江苏连云港222005
  • 相关基金:国家自然科学基金项目(50676063)、上海市重点学科(T0503;P0502)资助
中文摘要:

目的:测量PCR温度的准确性和均一性。方法:PCR主程序:变性95℃,30s,退火55℃,40s,延伸72℃,50s,6个循环;热电偶分别放在孔内和管内。结果:每个循环以及每次测量的孔和管内的温度重复性很好;变性阶段,孔的温度和管内液体温度与程序温度偏差较大,温度准确性都较差,但所有孔的温度都达到通常需要的变性温度(变性90℃以上),但是44.4%的孔在90℃以上停留的时间不能达到设置的时间;变性阶段,11.1%管内液体不能达到所需的变性温度(90℃).结论:这些缺陷可能导致扩增结果假阳性或假阴性。

英文摘要:

Both temperature accuracy of wells at block and uniformity of well-to-well and hatch-to batch were tested on a standard PCR process. Method: The primary thermal cycling settings were, 6 cycles of 95℃(denaturation)for 30 s,55 ℃( annealing )for 40 s,72 ℃ ( extension )for 50 s. Two thermal couples were embedded in bottom of wells and tubes respectively. Results: The temperatures of not only wells but also liquids in tubes measured had good repeatability cycle-to-cycle and batch-to-batch respectively. Temperatures of wells and in-tube liquids had poor uniformity during denaturation and deviated far from the programmed temperature. Although all wells reach the required denatured temperature (over 90℃),There are 44.4% wells which can not retain at all programmed times above 90℃ during denaturation.11.1% in-tube liquids cannot reach the common reaction temperature(over 90℃). Conclusion: False positive or false negative may be induced because of these shortcomings.

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期刊信息
  • 《生命科学仪器》
  • 主管单位:北京仪表集团有限责任公司
  • 主办单位:北京北分仪器技术公司
  • 主编:邓玉林
  • 地址:北京市丰台区星火路1号昌宁大厦四层4D
  • 邮编:100070
  • 邮箱:smkxyq@126.com
  • 电话:010-59495568 59492268 56035038
  • 国际标准刊号:ISSN:1671-7929
  • 国内统一刊号:ISSN:11-4846/TH
  • 邮发代号:2-262
  • 获奖情况:
  • 国内外数据库收录:
  • 被引量:2857