DETECTION OF SINGLE BASE MISMATCHES OF THYMINE AND CYTOSINE RESIDUES BY POTASSIUM-PERMANGANATE AND HYDROXYLAMINE IN THE PRESENCE OF TETRALKYLAMMONIUM SALTS
DETECTION OF SINGLE BASE MISMATCHES OF THYMINE AND CYTOSINE RESIDUES BY POTASSIUM-PERMANGANATE AND HYDROXYLAMINE IN THE PRESENCE OF TETRALKYLAMMONIUM SALTS
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DOI：
10.1093/nar/18.23.6807
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发表时间：
1990-12-11
影响因子：
14.9
通讯作者：
KAFATOS, FC
中科院分区：
文献类型：
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作者：
GOGOS, JA;KARAYIORGOU, M;KAFATOS, FC
在四甲基氯化铵存在下，高锰酸钾特异性修饰错配的胸腺嘧啶。 类似地，羟胺对错配胞嘧啶的修饰被四乙基氯化铵增强。 修饰后的哌啶裂解允许特异性鉴定T和C错配，并通过延伸，当分析相对的DNA链时，也可以鉴定A和G错配。 这些反应可以方便地进行与DNA固定在Hybond M-G纸。 我们描述了利用这些反应来检测错配的条件，例如使用合成寡核苷酸探针或特定基因组DNA序列的PCR扩增来检测点突变或遗传多态性。
In the presence of tetramethylammonium chloride, potassium permanganate specifically modifies mismatched thymines. Similarly, the modification of mismatched cytosines by hydroxylamine was enhanced by tetraethylammonium chloride. Modification followed by piperidine cleavage permits specific identification of the T and C mismatches and by extension, when the opposite DNA strand is analyzed, of A and G mismatches as well. These reactions can be performed conveniently with DNA immobilized on Hybond M-G paper. We describe conditions that exploit these reactions to detect mismatches, e.g. point mutations or genetic polymorphisms, using either synthetic oligonucleotide probes or PCR amplification of specific genomic DNA sequences.