Allen Angel G, Barthelson Karissa, Lardelli Michael
Alzheimer's Disease Genetics Laboratory, The University of Adelaide, Adelaide, SA 5005, Australia.
Biol Methods Protoc. 2023 Jul 31;8(1):bpad015. doi: 10.1093/biomethods/bpad015. eCollection 2023.
DNA size markers (also known as 'molecular weight markers' or 'DNA ladders') are an essential tool when using gel electrophoresis to identify and purify nucleic acids. However, the cost of these DNA ladders is not insignificant and, over time, impinges on the funds available for research and training in molecular biology. Here, we describe a method for the generation of 'pHAPE', a plasmid from which a variety of DNA ladders can be generated via simple restriction enzyme digestions. The pHAPE plasmid can be generated by mutagenesis of the commonly used pBluescript II SK+ phagemid followed by insertion of a 7141 bp sequence (comprised of three smaller, synthetic fragments). Our use of pHAPE allows us some small relief from the ever-rising costs of performing molecular biology experiments ('Don't worry, pHAPE').
DNA大小标记物（也称为“分子量标记物”或“DNA阶梯”）是在使用凝胶电泳鉴定和纯化核酸时必不可少的工具。然而，这些DNA阶梯的成本并非微不足道，随着时间的推移，会影响分子生物学研究和培训的可用资金。在这里，我们描述了一种生成“pHAPE”的方法，这是一种质粒，通过简单的限制性内切酶消化可以从中产生多种DNA阶梯。pHAPE质粒可以通过对常用的pBluescript II SK+噬菌粒进行诱变，然后插入一个7141 bp的序列（由三个较小的合成片段组成）来生成。我们对pHAPE的使用使我们在进行分子生物学实验不断上涨的成本方面得到了一些缓解（“别担心，有pHAPE”）。
