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两种微量DNA提取方法的比较 被引量：3

Comparison between two methods of DNA extraction
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摘要 目的:比较PromegaWizardGenomicDNA纯化试剂盒(PWGDPK)和国产星普液体微量DNA提取试剂盒(SLDPK)提取DNA的效果,探讨从冷冻保存的人外周血中提取基因组DNA的最佳方案。方法:采用反复冻融3次并在-20℃冻存1年的12例健康成人外周血样品,分别应用两种试剂盒提取基因组DNA,并采用吸收光谱法、琼脂糖凝胶电泳和PCR扩增等方法分别对提取的基因组DNA进行对比鉴定。结果:PWGDPK提取的DNA浓度为15.0～130.0mg·L-1[(74.8±39.3)mg·L-1],而SLDPK为15.5～37.0mg·L-1[(26.6±6.2)mg·L-1],前者显著高于后者(t=4.119,df=11,P<0.005)。但两种试剂盒提取的基因组DNA纯度都不高,大多数样品A260/A280比值偏离了1.8～2.0范围。PWGDPK提取的12个基因组DNA样品,琼脂糖凝胶电泳后都可见电泳带,9个条带较深,3个条带较浅。SLDPK提取的样品中,只有7个可见较浅的电泳带。应用两种试剂盒提取的12个基因组DNA样品作为模板进行PCR反应后,在rs1171558SNP位点均有11个样品(91.7%)获得目的片段;在rs2248745SNP位点,应用SLDPK只有4个样品(33.3%)获得目的片段,应用PWGDPK有10个样品(83.3%)获得目的片段,后者明显优于前者。结论:PWGDPK比SLDPK更适合于从长期冷冻保存且反复冻融过的全血中提取基因组DNA。
作者 张萱 王珍琦 史杰萍 赵景春 于雅琴 刘树铮 尉军
机构地区 吉林大学基因组医学研究中心卫生部放射生物学重点实验室
出处 《吉林大学学报（医学版）》 CAS CSCD 北大核心 2005年第4期636-639,共4页 Journal of Jilin University：Medicine Edition
基金 国家自然科学基金资助课题(30170343) 吉林大学创新基金课题
关键词 DNA/分离和提纯 聚合酶链反应 基因组
分类号 Q523 [生物学—生物化学]
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参考文献10

  • 1Twyman RM. SNP discovery and typing technologies for pharmacogenomics [J]. Curr Top Med Chem, 2004,4 (13):1423-1431.
  • 2Ceriotti G, Chessa S, Bolla P, et al. Single nucleotide polymorphisms in the ovine casein genes detected by polymerase chain reaction-single strand conformation polymorphism [J]. J Dairy Sci, 2004, 87 (8): 2606-2613.
  • 3Zhou GH, Shirakura H, Kamahori M, et al. A gel-free SNP genotyping method: bioluminometric assay coupled with modified primer extension reactions (BAMPER) directly from double-stranded PCR products [J]. Hum Mutat, 2004,24 (2) :155-163.
  • 4Dearlove AM. High throughput genotyping technologies [J].Brief Funct Genomic Proteomic, 2002, 1 (2): 139-150.
  • 5Che Y, Chen X. A multiplexing single nucleotide polymorphism typing method based on restriction-enzyme mediated single-base extension and capillary electrophoresis[J]. Anal Biochem, 2004, 329 (2): 220-229.
  • 6Bannai M, Higuchi K, Akesaka T, et al. Single-nucleotidepolymorphism genotyping for whole-genome amplified samples using automated fluorescence correlation spectroscopy [J]. Anal Biochem, 2004, 327 (2): 215-221.
  • 7孙祖玥,尉军,刘树铮.COMT基因多态性与精神分裂症的相关性[J].吉林大学学报（医学版）,2003,29(1):16-18. 被引量：1
  • 8郭英君,于雅琴,史杰萍,刘树铮,尉军.人类PIP3-E基因与精神分裂症的关系[J].吉林大学学报（医学版）,2004,30(2):163-165. 被引量：1
  • 9Zhang X, Wei J, Yu YQ, et al. Is NOTCH4 associated with schizophrenia [J]. Psychiatr Genet, 2004, 14 (1): 43-46.
  • 10颜子颖 王海林（译）.精编分子生物学实验指南[M].北京:科学出版社,1999.480-487.

二级参考文献14

  • 1[1]Pulver AE, Karayiorgou M, Wolyniec PS, et al. Sequential strategy to identify a susceptibility gene for schizophrenia: report of potenti al linkage on chromosome 22q12-q13.1: part 1[J]. Am J Med Genet, 1994, 54: 36-43.
  • 2[2]Ohmori O, Shinkai T, Kojima H, et al. Association study of a functional catechol-O-methyltransferase gene polymorphism in Japanese schizophren ics[J]. Neurosci Lett, 1998, 243(1-3): 109-112.
  • 3[3]Kunugi H, Vallada HP, Sham PC, et al. Catechol-O-methyltransfe rase polymorphisms and schizophrenia: a transmission disequilibrium study in multiply affected families[J]. Psychiatr Genet, 1997,7(3):97-101.
  • 4[4]Strous RD, Bark N, Parsia SS, et al. Analysis of a functional catechol-O-methyltransferase gene polymorphism in schizo-phrenia: evidence for association with aggressive and antisocial behavior[J]. Psychiatry Res, 1997, 69(2-3): 71-77.
  • 5[5]Daniels JK, Williams NM, Williams J, et al. No evidence for allelic association between schizophrenia and a polymorphism determining high or low catechol O-methyltransferase activity[J]. Am J Psychiatry, 1996, 153(2): 268 -270.
  • 6[6]Karayiorgou M, Gogos JA, Galke BL, et al. Identification of sequence variants and analysis of the role of the catechol-O-methyl-transferase gene in schizophrenia susceptibility[J]. Biol Psychiatry, 1998, 43(6): 425-431 .
  • 7Thaker GK, Carpenter WT Jr. Advances in schizophrenia [J].Nat Med, 2001, 7 (6): 667-671.
  • 8Bruzustowicz LM, Honer WG, Chow EW, et al. Use of aquantitive traittomapa locus associated with severity of positive symptoms in ilial schizophinia to chromosome 6p [J]. Am J Hum Gene, 1997, 61: 1388-1396.
  • 9Wright P, Donaldson PT, Underhill JA, et al. Genetic association of the HLADRB1 gene locus on chromosome 6p21.3 with schizophrenia [J]. Am J Psychiatry, 1996, 153: 1530-1533.
  • 10Daniels JK, Williams NM, Williams J, et al. No evidence for allelic association between schizophrenia and polymorphism determining high or low catechol O-methyl transferase activity [J]. Am J Psychiatry, 1996, 153 (2): 268-270.

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吉林大学学报（医学版）
2005年 第4期

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