3G vector-primer plasmid for constructing full-length-enriched cDNA libraries.
3G vector-primer plasmid for constructing full-length-enriched cDNA libraries.
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DOI：
10.1016/j.ab.2008.05.022
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发表时间：
2008-09
影响因子：
2.9
通讯作者：
Dong Zheng;Yanna Zhou;Zidong Zhang;Zai-yu Li;Xuedong Liu
中科院分区：
文献类型：
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作者：
Dong Zheng;Yanna Zhou;Zidong Zhang;Zai-yu Li;Xuedong Liu
我们设计了一个 3G 载体-引物质粒，用于生成全长富集互补 DNA (cDNA) 文库。该质粒（由聚dT末端和脱氧鸟苷[dG]末端组装而成）利用逆转录酶的末端转移酶活性和改良的链替换方法，将引发全长cDNA链合成和定向cDNA克隆结合起来。因此，减少了 cDNA 文库制备的步骤数量，同时简化了下游基因操作、测序和亚克隆。 3G 载体-引物质粒方法产生完全代表性的质粒引物文库，与 SMART（RNA 转录物 5' 端的转换机制）方法产生的文库等效。
We designed a 3G vector–primer plasmid for the generation of full-length-enriched complementary DNA (cDNA) libraries. By employing the terminal transferase activity of reverse transcriptase and the modified strand replacement method, this plasmid (assembled with a polydT end and a deoxyguanosine [dG] end) combines priming full-length cDNA strand synthesis and directional cDNA cloning. As a result, the number of steps involved in cDNA library preparation is decreased while simplifying downstream gene manipulation, sequencing, and subcloning. The 3G vector–primer plasmid method yields fully represented plasmid primed libraries that are equivalent to those made by the SMART (switching mechanism at 5′ end of RNA transcript) approach.