） 摘要： 目的：探讨运用PCR方法快速筛选DNA文库的可行性
PCR可以应用于快速筛选DNA文库
荧光原位杂交证实该基因位于染色体1q2
10-q2
11
采用噬菌斑原位杂交和PCR相结合的方法
对猪的gDNA文库进行α-1,3GT基因筛选
方法 以猪α-1,3GT cDNA片段为探针
） Abstract： Objective: To explore the feasibility of quickly screening of gDNA library with PCR technique
Methods: We adopted porcine α-1,3GT cDNA fragment as the probe, used the primers synthesized by the specific sequence on cDNA to carry out α-1,3GT gene screening of porcine gDNA library by combining PCR and in situ plaque hybridization, and then performed enzymic digestion, southern blot, sequencing and fluorescence in situ hybridization for location
Results: After having finished one-time hybridization and one-time PCR, we obtained 7 positive monoclones with very strong signals, and each insert length of them is over 8kb, including the third intron
Moreover, 3 tested clones among them contain the third and fourth exons according to the sequencing results, and FISH mapped the inserts of the 3 clones to pig chromosome 1q2
10-q2
11
Conclusion: PCR could be applicable to the quick screening of DNA library and much simpler than the conventional in situ plaque hybridization only used
Keywords： genomic DNA library, PCR, hybridization 下载PDF阅读器 PDF全文下载： 初稿 （ 588 ） 作者简介： 通信联系人： 【收录情况】 中国科技论文在线： 赵永祥
PCR技术在快速筛选DNA文库中的应用[EB/OL]
北京：中国科技论文在线
经酶切、Southern Blot、测序和荧光原位杂交定位
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