Rational primer design greatly improves differential display-PCR (DD-PCR).
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.
Graf
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Lymphocyte Development Group, MRC Clinical Sciences Centre, Royal Postgraduate Medical School, Hammersmith Hospital, Du Cane Road, London W12 0NN, UK. dgraf@rpms.ac.uk
Fisher
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Merkenschlager
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D000818:Animals; D002417:Cattle; D017931:DNA Primers / Q000138:chemical synthesis*; D015195:Drug Design; D016133:Polymerase Chain Reaction / Q000379:methods*; D013950:Thymus Gland / Q000166:cytology
D017931:DNA Primers