Isolation of differentially expressed genes by combining representational difference analysis (RDA) and cDNA library arrays.

نویسندگان

  • M Geng
  • C Wallrapp
  • F Müller-Pillasch
  • M Frohme
  • J D Hoheisel
  • T M Gress
چکیده

The difference products (DP) of representational difference analyses (RDA) were used as hybridization probes on cDNA arrays. The effectivity of RDA products obtained with increasing driver/tester ratios (DP 1 = 100:1, DP 2 = 800:1 and DP 3 = 400,000:1) to isolate differentially expressed genes was compared with the effectivity of conventional differential hybridizations. Pacreatic cancer and control tissues were used as a test system to isolate differentially expressed genes. The use of RDA products as hybridization probes showed two major advantages: (i) a reliable identification of true differential signals; and (ii) only one autoradiograph had to be analyzed, which eliminated the need for a laborious subtraction of signal intensities obtained with different cDNA probes. Increasing driver/tester ratios in iterative rounds of RDA delivered more specific results, though the total yield of differential clones was gradually reduced. In this situation, the intermediate RDA product DP 2 provided the best compromise.

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عنوان ژورنال:
  • BioTechniques

دوره 25 3  شماره 

صفحات  -

تاریخ انتشار 1998