方法：使用聚合酶链式反应（PCR）简化DNA梯状条带的制备

Methodology: simplified preparation of a DNA ladder using PCR.

作者信息

Wang T-Y, Wang L, Wang F

机构信息

Department of Biochemistry and Molecular Biology, Xinxiang Medical University, Henan, P.R. China.

出版信息

Genet Mol Res. 2011;10(3):1631-5. doi: 10.4238/vol10-3gmr1177.

DOI:10.4238/vol10-3gmr1177

PMID:21863555

Abstract

Serving as a DNA molecular weight standard, the DNA ladder has been widely used in molecular biology applications. We developed a simple method for the preparation of a DNA marker, which involves designing primers to amplify 100- to 1000-bp DNA fragments using lambda DNA as a template for polymerase chain reaction, followed by extraction with phenol/chloroform, precipitation with ethanol and mixing. Fragments of 100- to 1000-bp DNA were successfully amplified; the sequences showed 100% identity with lambda DNA. This prepared DNA marker displayed clear bands, indicating that it can be used for molecular studies.

摘要

DNA 梯度标准作为一种 DNA 分子量标准，已广泛应用于分子生物学领域。我们开发了一种制备 DNA 标记物的简单方法，该方法包括设计引物，以λDNA 为模板通过聚合酶链式反应扩增 100 至 1000 碱基对的 DNA 片段，随后用苯酚/氯仿萃取，乙醇沉淀并混合。成功扩增出了 100 至 1000 碱基对的 DNA 片段；这些序列与λDNA 显示出 100%的同一性。这种制备的 DNA 标记物显示出清晰的条带，表明它可用于分子研究。