| 小麦cDNA文库构建方法的改进及优化 |
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| 引用本文： | 许延芳,潘丽娜,刘晓颖,王振英. 小麦cDNA文库构建方法的改进及优化[J]. 天津农业科学, 2012, 18(3): 22-25 |
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| 作者姓名： | 许延芳 潘丽娜 刘晓颖 王振英 |
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| 作者单位： | 天津师范大学生命科学学院,天津,300387 |
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| 基金项目： | 天津师范大学博士基金，国家自然科学基金 |
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| 摘 要： | 基于SMART法,优化cDNA文库构建方法。将常规SMART法中的限制性内切酶SfiI替换成BsaI,在引物两侧分别加入BsaI-XhoI和BsaI-BamHI酶切位点,仅使用BsaI单酶切即可获得两不同的粘性末端,可用于连接任意载体。应用此方法构建了小麦cDNA文库,获得2.7×104个独立克隆,文库平均长度500bp。
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| 关 键 词： | cDNA文库 SMART法 BsaI |
An Improved, Efficient Method for cDNA Library Construction of Triticum aestivum |
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| XU Yan-fang , PAN Li-na , LIU Xiao-ying , WANG Zhen-ying. An Improved, Efficient Method for cDNA Library Construction of Triticum aestivum[J]. Tianjin Agricultural Sciences, 2012, 18(3): 22-25 |
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| Authors: | XU Yan-fang PAN Li-na LIU Xiao-ying WANG Zhen-ying |
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| Affiliation: | (College of Life Science, Tianjin Normal University, Tianjin 300387, China) |
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| Abstract: | A low-cost method based on SMART technique for cDNA library construction in Triticum aestivum was provied. In this study, the restriction endonuclease SfiI in the SMART technique was changed by BsaI, and the primer with BsaI-XhoI or BsaI-BamHI binding site was used for double-strand cDNA enrichment. The double-strand cDNA was digested by restriction endonuclease BsaI, which can form two different cohesive termini, and then it can be connected with any vectors. As a result, more than 2.7×104 independent clones were collected in the cDNA library, and the average sequence of inserts was 500 bp. |
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| Keywords: | cDNA library SMART method BsaI |
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