腺病毒DNA的羟基磷灰石层析与甲酰胺变性
Hydroxyapatite chromatography and formamide denaturation of adenovirus DNA.
作者信息
Tibbetts C, Johansson K, Philipson L
出版信息
J Virol. 1973 Aug;12(2):218-25. doi: 10.1128/JVI.12.2.218-225.1973.
Abstract
Denatured adenovirus DNA was retained by hydroxyapatite columns under conditions generally used for selective retention of double-stranded DNA, probably due to several partially complementary sequences within single-stranded DNA. It was found that addition of formamide reduced the fraction of sonically treated, denatured adenovirus DNA bound to hydroxyapatite from about 30% to less than 1%. This led to a study of the effect of formamide on the melting temperature (T(m)) of double-stranded DNA in solution or bound to hydroxyapatite. The T(m) of DNA decreases 0.56 C/1% formamide, a value determined in buffered solutions with purified formamide.
摘要
在通常用于选择性保留双链DNA的条件下，变性腺病毒DNA被羟基磷灰石柱保留，这可能是由于单链DNA内存在几个部分互补序列。研究发现，添加甲酰胺可使经超声处理的变性腺病毒DNA与羟基磷灰石结合的比例从约30%降至1%以下。这引发了对甲酰胺对溶液中或与羟基磷灰石结合的双链DNA解链温度（T(m)）影响的研究。DNA的T(m)每1%甲酰胺降低0.56℃，该值是在含有纯化甲酰胺的缓冲溶液中测定的。
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