摘要

S1脉冲场凝胶电泳（PFGE）是一种从质粒核酸中分离细菌染色体的方法。 当与溴化乙锭染色和UV可视化相结合时，该方法在评估单个细菌菌株中质粒的数量方面非常出色。 当针对DNA分子标记运行时，它也很好地逼近每个单独质粒的真实大小。 然而，下游应用如：各个质粒或染色体上个体抗性基因的位置受到大DNA分子从琼脂糖凝胶到吸附尼龙或硝酸纤维素膜的非常差的转移的阻碍。 在此，我们描述了一种直接探测琼脂糖PFGE凝胶的方法，消除了转移的必要性并产生出色的基因组定位结果。

S1 pulsed field gel electrophoresis (PFGE) is a method to separate the bacterial chromosome(s) from plasmid nucleic acids. When combined with ethidium bromide staining and UV visualization this method is excellent at assessing the number of plasmids in individual bacterial strains. It is also good at approximating the true size of each individual plasmid when run against a DNA molecular marker. However, downstream applications such as: the location of individual resistance genes on individual plasmids or the chromosome are hampered by very poor transfer of large DNA molecules from agarose gels to adsorbent nylon or nitrocellulose membranes. Herein, we describe a method to directly probe agarose PFGE gels eliminating the necessity of transfer and generating excellent genomic location results.

https://link.springer.com/protocol/10.1007/978-1-4939-7638-6_12