用银染法测定人睫状神经营养因子基因的核苷酸序列  

DNA SEQUENCING OF HUMAN CILLIARY NEUROTROPIC FACTOR GENE BY SILVER STAINING METHOD

作　　者：胡向阳[1] 黄晓明[1] 高翼之[1] 

机构地区：[1]南京铁道医学院科研所分子生物学研究室

出　　处：《中华医学遗传学杂志》1998年第1期49-50,共2页Chinese Journal of Medical Genetics

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。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.

关 键 词：DNA序列分析 银染法 hCNTF 核苷酸 

分 类 号：Q75[生物学—分子生物学]