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用银染法测定人睫状神经营养因子基因的核苷酸序列
引用本文:胡向阳,黄晓明,高翼之. 用银染法测定人睫状神经营养因子基因的核苷酸序列[J]. 中华医学遗传学杂志, 1998, 15(1): 49-50
作者姓名:胡向阳  黄晓明  高翼之
作者单位:南京铁道医学院科研所分子生物学研究室
摘    要:目的介绍一种采用银染法的非同位素DNA序列分析技术。方法用TaqDNA聚合酶扩增DNA模板进行测序反应,在测序反应中不使用任何标记,测序凝胶上的DNA条带直接用灵敏的银染法检出。结果用银染法测定了人睫状神经营养因子基因的核苷酸序列,得到的DNA测序图谱与同位素相近,但全部结果可在测序反应后2小时内得到。结论银染法是一种十分经济方便并且快速可行的DNA测序方法。

关 键 词:DNA序列分析  银染法  人睫状神经营养因子

DNA SEQUENCING OF HUMAN CILLIARY NEUROTROPIC FACTOR GENE BY SILVER STAINING METHOD
Hu Xiangyang ,Huang Xiaoming,Gao Yizhi. DNA SEQUENCING OF HUMAN CILLIARY NEUROTROPIC FACTOR GENE BY SILVER STAINING METHOD[J]. Chinese journal of medical genetics, 1998, 15(1): 49-50
Authors:Hu Xiangyang   Huang Xiaoming  Gao Yizhi
Affiliation:Hu Xiangyang *,Huang Xiaoming,Gao Yizhi.*Institute of Medical Sciences,Nanjing Railway Medical College,Nanjing,210009
Abstract:Objective To introduce a non radioactive protocol for DNA sequence analysis which employs a silver staining procedure Methods DNA template is amplified by Taq DNA polymerase to generate sequence ladder,No labeled nucleotide or primer is involved in the thermal cycling sequencing reaction.A sensitive silver staining procedure is employed to visualize bands in sequencing gel.Results This silver staining method is used for DNA sequence analysis of human cilliary neurotrophic factor(hCNTF) gene.The resulted band resolution is comparable to radioactive sequencing method,and data can be obtained within less than 2 hours after sequencing reaction.Conclusion Silver staining is a rapid,inexpensive and reliable method for DNA sequence analysis.
Keywords:DNA sequence analysis Silver staining Human cilliary neurotropic factor
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