Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1997 Jun 1;25(11):2239–2240. doi: 10.1093/nar/25.11.2239

Rational primer design greatly improves differential display-PCR (DD-PCR).

D Graf 1, A G Fisher 1, M Merkenschlager 1
PMCID: PMC146700  PMID: 9153330

Abstract

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.

Full Text

The Full Text of this article is available as a PDF (80.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bauer D., Müller H., Reich J., Riedel H., Ahrenkiel V., Warthoe P., Strauss M. Identification of differentially expressed mRNA species by an improved display technique (DDRT-PCR). Nucleic Acids Res. 1993 Sep 11;21(18):4272–4280. doi: 10.1093/nar/21.18.4272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Liang P., Averboukh L., Pardee A. B. Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization. Nucleic Acids Res. 1993 Jul 11;21(14):3269–3275. doi: 10.1093/nar/21.14.3269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Liang P., Pardee A. B. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science. 1992 Aug 14;257(5072):967–971. doi: 10.1126/science.1354393. [DOI] [PubMed] [Google Scholar]
  4. Mages H. W., Stamminger T., Rilke O., Bravo R., Kroczek R. A. Expression of PILOT, a putative transcription factor, requires two signals and is cyclosporin A sensitive in T cells. Int Immunol. 1993 Jan;5(1):63–70. doi: 10.1093/intimm/5.1.63. [DOI] [PubMed] [Google Scholar]
  5. Marchuk D., Drumm M., Saulino A., Collins F. S. Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucleic Acids Res. 1991 Mar 11;19(5):1154–1154. doi: 10.1093/nar/19.5.1154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Sehgal A., Presente A., Dudus L., Engelhardt J. F. Isolation of differentially expressed cDNAs during ferret tracheal development: application of differential display PCR. Exp Lung Res. 1996 Jul-Aug;22(4):419–434. doi: 10.3109/01902149609046033. [DOI] [PubMed] [Google Scholar]
  7. Zipfel P. F., Irving S. G., Kelly K., Siebenlist U. Complexity of the primary genetic response to mitogenic activation of human T cells. Mol Cell Biol. 1989 Mar;9(3):1041–1048. doi: 10.1128/mcb.9.3.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES