WO2019228542A9 - 引物5'端反向互补的荧光pcr及试剂盒 - Google Patents

引物5'端反向互补的荧光pcr及试剂盒 Download PDF

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Publication number
WO2019228542A9
WO2019228542A9 PCT/CN2019/094329 CN2019094329W WO2019228542A9 WO 2019228542 A9 WO2019228542 A9 WO 2019228542A9 CN 2019094329 W CN2019094329 W CN 2019094329W WO 2019228542 A9 WO2019228542 A9 WO 2019228542A9
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primer
amplification
reverse complementary
product
fluorescent pcr
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PCT/CN2019/094329
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English (en)
French (fr)
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WO2019228542A1 (zh
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江洪
江和来
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江洪
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Publication of WO2019228542A1 publication Critical patent/WO2019228542A1/zh
Publication of WO2019228542A9 publication Critical patent/WO2019228542A9/zh

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

本申请公开一种引物5'端反向互补的荧光PCR，其特征是一对引物5'端添加一段序列反向互补的荧光PCR扩增，带来扩增产物间末端互补，靶产物3'末端也可以互为模板、互为引物而增加扩增效率，在常规PCR指数放大10 9-12基础上再增加灵敏度50倍，或同样灵敏度情况下可少扩增5个循环；另一面引物5'端互补不会延伸反而抑制原引物对3'末端间的聚合非特异性或少扩增5个循环可避开引物非特异扩增区。联合中部同序与3'最末端倒数第1-2个碱基A腺嘌呤结尾的引物+UDG酶解dU代替dT的扩增产物,和矿物油密闭使PCR反应循环内无引物二聚体PD产生及杜绝产物气雾胶交叉污染。
PCT/CN2019/094329 2018-05-31 2019-07-02 引物5'端反向互补的荧光pcr及试剂盒 WO2019228542A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810544265.6A CN108796047A (zh) 2018-05-31 2018-05-31 引物5’端反向互补的荧光pcr
CN201810544265.6 2018-05-31

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WO2019228542A1 WO2019228542A1 (zh) 2019-12-05
WO2019228542A9 true WO2019228542A9 (zh) 2020-01-09

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108796047A (zh) * 2018-05-31 2018-11-13 江洪 引物5’端反向互补的荧光pcr
CN111662962B (zh) * 2020-06-09 2023-06-23 珠海市坤元科技有限公司 一种双向链替换环循环的核酸恒温扩增法
CN113832147B (zh) * 2021-09-08 2024-06-14 华南农业大学 一种高效的大片段dna合成与扩增的pcr引物、方法及应用

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020197611A1 (en) * 2001-06-21 2002-12-26 Chagovetz Alexander Michael Method for real-time detection and quantification of nucleic acid sequences using fluorescent primers
CN102146432B (zh) * 2010-02-04 2017-02-15 北京泰格瑞分子检验有限公司 一对部分同序引物减少其二聚体的方法
CN102352350A (zh) * 2011-09-30 2012-02-15 北京万达因生物医学技术有限责任公司 一种同序引物置换的核酸扩增技术
CN103114131B (zh) * 2012-11-30 2018-10-02 珠海市坤元农业科技有限公司 一种引物中部序列干扰pcr技术
CN106191214B (zh) * 2015-04-30 2019-05-10 奥健生物科技（广州）有限公司 一种多色荧光熔解曲线pcr检测方法
CN108085372B (zh) * 2018-01-02 2021-09-28 江洪 引物末端酶解二聚体法pcr
CN108796047A (zh) * 2018-05-31 2018-11-13 江洪 引物5’端反向互补的荧光pcr

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WO2019228542A1 (zh) 2019-12-05

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