Ledakis P, Tanimura H, Fojo T
Medicine Branch, National Cancer Institute, Bldg. 10, Rm. 12N226, 9000 Rockville Pike, Bethesda, Maryland, 20892, USA.
Biochem Biophys Res Commun. 1998 Oct 20;251(2):653-6. doi: 10.1006/bbrc.1998.9517.
Since its original description, differential display PCR (DD-PCR) has been extensively used in attempts to identify novel genes under a variety of circumstances. Despite its widespread use, however, few novel genes of interest have been identified. In the present study we describe a set of experiments examining reasons for failure of differential display. Evidence is presented that aberrant priming at both the 5' and 3' ends results in competition in the PCR, precluding detection of messages other than those which are abundantly expressed. Appropriate calculations are discussed which indicate this was predictable and unlikely to be overcome. While DD may be successfully applied in some settings, the evidence indicates that only abundantly expressed messages can be detected. This limitation is emphasized.
自从差异显示PCR（DD-PCR）最初被描述以来，它已被广泛用于在各种情况下鉴定新基因的尝试中。然而，尽管它被广泛使用，但很少有感兴趣的新基因被鉴定出来。在本研究中，我们描述了一组实验，研究差异显示失败的原因。有证据表明，5'和3'末端的异常引物引发导致PCR中的竞争，排除了除大量表达的信息之外的其他信息的检测。讨论了适当的计算方法，这些计算表明这是可预测的，并且不太可能被克服。虽然DD可能在某些情况下成功应用，但证据表明只能检测到大量表达的信息。强调了这一局限性。
