Subtractive Hybridization for the Identification of Differentially Expressed Genes Using Uraci-DNA Glycosylase and Mung-Bean Nuclease

Abstract

Subtractive hybridization is a technique for detecting differences of gene expression between two populations of RNAs or complementary DNAs (cDNAs). It is based on base pair complementary that nucleic acid sequences in common with the two populations can form hybrids. After hybridization, the hybrids are removed and the sequences in only one population are identified. This technique can be used for detecting differences between the mRNA in different cells, tissues, and organisms, or cells under two different conditions, tissues of various stages of development and growth, and treatments of hormone or other modulators. One of the common uses of subtractive hybridization is for the detection of DNA differences between different genomes or between cell types to identify certain types of genomic rearrangement as well as differentially expressed genes. For example, a recently developed method of subtractive hybridization has been used to identify cDNAs associated with activin-mediated inhibition of cell growth in human prostate cancer cells (1). In addition to the detection of genes differentially expressed in the same cell type under two different conditions, this technique can be used to ascertain the quantity of mRNAs and cDNAs generated (2).