从凝胶中回收银染DNA条带的研究
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Q523.8 Q785

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广东省自然科学基金,( 960 14 9),卫生部优秀青年人才基金 ( 970 16)


Study on the Silver-staining DNA bands recovered from polyacrylamide gels
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    摘要:

    目的:比较常规克隆测序与从凝胶中回收银染DNA条带进行直接测序和克隆测序的特点。方法:用单链构像多态法分离杂合性突变DNA样品,然后从凝胶中回收变异的银染DNA条带进行直接测序和克隆测序,并与常规克隆测序进行比较。结果:从凝胶中回收DNA片段直接测序的正确率超过97%,回收片段克隆测序的正确率与常规克隆测序相同,达100%。结论:从凝胶中回收DNA片段直接测序和克隆测序较常规克隆测序简便实用,在基因克隆,突变鉴定等工作中有一定实用价值。

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    Objective To compare the feasibility,reliability and accuracy of cloning sequencing and direct sequencing for DNA fragments recovered from polyacrylamide gels after silver staining with that of routine cloning sequencing.Methods Heterozygous mutant DNA sample was separated by using single strand conformational polymorphism(SSCP)analysis.The mutant DNA fragments,recovered from polyacrylamide gels after silver staining,were analyzed by cloning sequencing and direct sequencing.The DNA sequence was compared with that from routine cloning sequencing.Results The correctness rate of direct sequencing for recovered DNA is above 97% and that of cloning sequencing is 100% which is the same as that of routine cloning sequencing.Conclusion Sequencing of specific DNA fragments recovered from gels is a rapid,practical and reliable way and is of usefulness in gene cloning and mutational identification.

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肖学珊,张清炯,黎仕强,贾小云,郭向明,邝志和,申惶煊.从凝胶中回收银染DNA条带的研究[J].临床检验杂志,2002,(1):24-27

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