RLCS, Restriction Landmark cDNA Scanning
It is important to identify and isolate differentially expressed genes whose expression underlies many biological processes such as development, differentiation, and cellular response to various stimuli, and also pathological changes that arise in diseases. Classically, two hybridization methods, differential and subtractive hybridization, have been used to analyze and isolate such genes [1–3]. However, differential hybridization is effective only for mRNAs expressed abundantly in one of the two types of cells. Subtractive hybridization is rather empirical and poor in reproducibility. Furthermore, both methods are time-consuming. Recently, Liang and Pardee have developed a novel method called differential display using an arbitrarily primed reverse transcription-coupled PCR and sequencing gels . This has proved to be a powerful technique for detecting changes in eukaryotic gene expression; it is simple, sensitive, and time-saving. However, PCR-mediated amplification with arbitrary primers requires delicate control, depending on the primers used [5,6], and cDNA probes thus obtained often give false positive signals on Northern blot analysis [7,8]. Furthermore, the differential display seems less sensitive in detecting rare mRNA populations, since it has a strong bias toward higher abundance mRNAs .
KeywordsDifferential Display mRNA Species Anchor Primer Restriction Landmark Genomic Scanning Rare mRNA
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