Reciprocal Subtraction Differential RNA Display (RSDD)
Identification of differentially expressed genes is an essential step in comprehending the molecular basis of complex physiological and pathological processes. Subtraction hybridization and differential RNA display (DDRT-PCR) are two methods that are widely and successfully employed to clone differentially expressed genes. Unfortunately, both methods have inherent problems and limitations requiring improvements in the technique. A combination of these two methods termed reciprocal subtraction differential RNA display is described here that considerably reduces the complexity of DDRT-PCR and facilitates the rapid and efficient identification and cloning of both abundant and rare differentially expressed genes.
Key WordsCloning differential RNA display reciprocal subtraction differential RNA display RSDD subtraction hybridization tumor progression gene expression
- 15.Jiang, H. and Fisher, P. B. (1993) Use of a sensitive and efficient subtraction hybridization protocol for the identification of genes differentially regulated during the induction of differentiation in human melanoma cells. Mol. Cell Differ. 1, 285–299.Google Scholar
- 19.Hay, B. and Short, J. M. (1992) ExAssist™ helper phage and SOLR™ for lambda ZAP II excisions. Strategies 5, 16–18.Google Scholar
- 20.Sambrook, J. and Russell, D. W. (ed.) (2001) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Lab. Press, Cold Spring Harbor, NY.Google Scholar