Gopalakrishnan R, Joseph S, Sellappa S
Department of Biotechnology, School of Life Sciences, Karpagam University, Coimbatore, Tamil Nadu, India.
Iran J Microbiol. 2010 Dec;2(4):210-2.
DNA ladder contains DNA fragments of different length but with known size, used to determine the size of unknown DNA molecules. Different DNA ladders are available for expected DNA length. Conserved sequences were selected for design of primers to generate DNA fragments of known specific size.
In this study, we describe a method by which DNA ladder was prepared based on multiplex PCR technique. Different lengths of DNA fragments were amplified using the primers designed according to the 1216-2136 sequence extent of lambda phage DNA. Target DNA fragments were amplified using multiplex PCR and extracted.
The results showed an amplified lambda phage DNA at particular target sites by using 1 forward and 6 different reverse primers (for 100, 200, 400, 600, 800, 1000bp) for the successful amplification.
This method would be more cost effective than commercial DNA molecular weight markers.
DNA阶梯包含不同长度但大小已知的DNA片段，用于确定未知DNA分子的大小。针对预期的DNA长度有不同的DNA阶梯可供选择。选择保守序列来设计引物，以产生已知特定大小的DNA片段。
在本研究中，我们描述了一种基于多重PCR技术制备DNA阶梯的方法。使用根据λ噬菌体DNA的1216 - 2136序列范围设计的引物扩增不同长度的DNA片段。通过多重PCR扩增并提取目标DNA片段。
结果显示，使用1条正向引物和6条不同的反向引物（用于100、200、400、600、800、1000bp）在特定目标位点成功扩增出λ噬菌体DNA。
该方法比商业DNA分子量标准更具成本效益。
