OPTIMAL CONDITIONS FOR SUPERCOIL DNA SEQUENCING WITH THE ESCHERICHIA-COLI DNA-POLYMERASE-I LARGE FRAGMENT
OPTIMAL CONDITIONS FOR SUPERCOIL DNA SEQUENCING WITH THE ESCHERICHIA-COLI DNA-POLYMERASE-I LARGE FRAGMENT
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DOI：
10.1016/0735-0651(88)90024-6
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发表时间：
1988-03-01
期刊：
影响因子：
--
通讯作者：
PENE, JJ
中科院分区：
文献类型：
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作者：
LIM, HM;PENE, JJ
用大肠杆菌DNA聚合酶Klenow片段从超螺旋DNA模板产生的测序梯通常由于高背景和错误掺入的核苷酸而不可读。这项研究表明，污染的RNA分子可以干扰模板：引物杂交。提供了用于纯化模板DNA的程序和克服这些问题的引物-模板杂交的严格条件。
Sequencing ladders produced from supercoiled DNA templates with the Escherichia coli DNA polymerase Klenow fragment are often unreadable because of a high background and misincorporated nucleotides. This study showed that contaminating RNA molecules can interfere with template:primer hybridization. Procedures are provided for the purification of template DNA and stringent conditions for primer-template hybridization that overcome these problems.