Graf D, Fisher A G, Merkenschlager M
Lymphocyte Development Group, MRC Clinical Sciences Centre, Royal Postgraduate Medical School, Hammersmith Hospital, Du Cane Road, London W12 0NN, UK.
Nucleic Acids Res. 1997 Jun 1;25(11):2239-40. doi: 10.1093/nar/25.11.2239.
Since its conception in 1992, differential display PCR (DD-PCR) has attracted widespread interest. Theoretically an attractive cloning approach, it combines the comparative analysis of several samples with the sensitivity of PCR. Although a large number of studies embracing this technology have been initiated, few novel genes of interest have been identified, suggesting that the method has not realised its potential. The present report shows that by modifying primer design, sampling of differentially expressed genes can be greatly enhanced and relevant genes can be isolated. Using our modified conditions DD-PCR efficiently screens a wide range of gene expression levels, in which differences are represented on a linear scale.
自1992年差异显示PCR(DD-PCR)概念提出以来,它已引起广泛关注。从理论上讲,它是一种有吸引力的克隆方法,将几个样本的比较分析与PCR的灵敏度结合在一起。尽管已经开展了大量采用该技术的研究,但很少鉴定出感兴趣的新基因,这表明该方法尚未发挥其潜力。本报告表明,通过修改引物设计,可以大大增强差异表达基因的采样,并分离出相关基因。使用我们修改后的条件,DD-PCR能够有效筛选广泛的基因表达水平,其中差异以线性比例呈现。