Construction of a circular single-stranded DNA template containing a defined lesion
Construction of a circular single-stranded DNA template containing a defined lesion
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DOI：
10.1016/j.dnarep.2009.03.006
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发表时间：
2009-07-04
期刊：
影响因子：
3.8
通讯作者：
Woodgate, Roger
中科院分区：
文献类型：
--
作者：
Karata, Kiyonobu;Vidal, Antonio E.;Woodgate, Roger
We report a concise and efficient method to make a circular single-stranded DNA containing a defined DNA lesion. In this protocol, phagemid DNA containing Uracil is used as a template to synthesize a complementary DNA strand using T7 DNA polymerase and an oligonucleotide primer including a site-specific DNA lesion. The ligated lesion-containing strand can be recovered after the phage-derived template DNA is degraded by treatment with E coli Uracil DNA glycosylase and Exonucleases I and III. The resulting product is a circular single-stranded DNA containing a defined DNA lesion suitable for in vitro translesion replication assays. Published by Elsevier B.V.
