Screening differentially expressed genes from suppression subtractive hybridizational positive clones using reverse mRNA dot blot technique

Purpose: To establish a method for screening the differential expression genes from the suppression subtractive hybridization positive clones. Methods: The PCR products of suppression subtractive hybridization positive clones were arrayed onto nylon membrane, hybridized with a labelled polyA {++RNA} probe and investigated with chemical luminescence reagent. The intensity of the dots was scanned and analyzed using Leica Q550Iw image analysis system. The intensity of the dots were calculated as average intensity per unit area which eliminates the intensity of background. If the ratio (R) of intensity of a pair of dots in two parallel arrays is ≥2.00, it is defined to be positive differential expression gene. Results: The differential expression genes are efficaciously screened from suppression subtractive hybridization positive clones using reverse mRNA dot blot technique. Conclusion: The reverse mRNA dot blot is a useful method for screening differential expression genes from suppression subtractive hybridization positive clones.