Detection of Modified Bases in Bacteriophage Genomic DNA
Detection of Modified Bases in Bacteriophage Genomic DNA
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DOI：
10.1007/978-1-0716-0876-0_5
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发表时间：
2021-01-01
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影响因子：
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通讯作者：
Weigele, Peter R.
中科院分区：
文献类型：
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作者：
Lee, Yan-Jiun;Weigele, Peter R.
总的来说，dsDNA尾噬菌体（尾病毒目）含有迄今为止观察到的DNA中天然存在的脱氧核苷酸的最大化学多样性。不断发现的新的修饰表明还有更多的修饰有待发现，因此，观察和表征非规范核苷的方法是及时的。我们在这里提出的协议提取基因组DNA从噬菌体颗粒，酶水解的DNA游离核苷，并检查核苷组合物的HPLC和质谱。
Collectively, the dsDNA tailed bacteriophages (Caudovirales) contain the largest chemical diversity of naturally occurring deoxynucleotides in DNA observed to date. The continuing discovery of new modifications in phages suggest many more are waiting to be found. Thus, methods for the observation and characterization of noncanonical nucleosides are timely. We present here protocols for extraction of genomic DNA from bacteriophage particles, enzymatic hydrolysis of DNA to free nucleosides, and examination of nucleoside composition by HPLC and mass spectrometry.