A simple and efficient method to reduce nontemplated nucleotide addition at the 3′ terminus of RNAs transcribed by T7 RNA polymerase
A simple and efficient method to reduce nontemplated nucleotide addition at the 3′ terminus of RNAs transcribed by T7 RNA polymerase
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DOI：
10.1017/s1355838299991033
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发表时间：
1999-09-01
期刊：
影响因子：
4.5
通讯作者：
Rüdisser, S
中科院分区：
文献类型：
--
作者：
Kao, C;Zheng, M;Rüdisser, S
在 5' 末端的最后两个核苷酸处用 C2'-甲氧基修饰的 DNA 模板显着减少了 T7 RNA 聚合酶从单链和双链 DNA 模板中添加非模板核苷酸。该策略用于生成几种不同的转录本。核磁共振波谱证明其中两个转录本的序列不受影响；从修改后的模板产生的转录本可以更容易地纯化，并且应该在许多应用中有用。
DNA templates modified with C2'-methoxyls at the last two nucleotides of the 5' termini dramatically reduced nontemplated nucleotide addition by the T7 RNA polymerase from both single- and double-stranded DNA templates. This strategy was used to generate several different transcripts. Two of the transcripts were demonstrated by nuclear magnetic resonance spectroscopy to be unaffected in their sequence; Transcripts produced from the modified templates can be purified with greater ease and should,be useful in a number of applications.