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随着cDNA文厍应用的日益广泛,其构建方法也有了很大改进和提高。获得完整和均一的mRNA是构建成功的基础,合成cDNA时可根据不同目的选择相应的反转录酶和方法,用于构建的载体也由质粒发展到噬菌粒并各有其利弊,双链cDNA克隆前应视连接方案进行末端修饰和分级分离,克隆时则需把握cDNA和载体的比例。近来发展起来的PCR介导的cDNA文库、消减cDNA文库、标准化cDNA文序和染色体或区域特异性cDNA文库进一步满足了不同的分子生物学需要。
With the increasing use of cDNA literatures, its construction methods have also been greatly improved and improved. Obtaining complete and homogeneous mRNAs is the basis of successful construction. The corresponding reverse transcriptase and the method can be selected according to different purposes when synthesizing cDNA. The vectors used for the construction also develop from plasmids to phagemids, Before cDNA cloning, the terminal modification and fractionation should be performed according to the ligation scheme. When cloning, it is necessary to determine the ratio of cDNA to vector. Recently developed PCR-mediated cDNA libraries, subtractive cDNA libraries, standardized cDNA sequences and chromosomal or region-specific cDNA libraries further satisfy different molecular biology needs.