Enhanced concatemer cloning - a modification to the SAGE (Serial Analysisof Gene Expression) technique

摘要

The Serial Analysis of Gene Expression (SAGE) method, described in 1995 by Velculescu et al,, represents a powerful means to compare gene expression between two mRNA populations. An improvement to SAGE that removes contaminating linker molecules, which compromise the efficiency of the method, has been developed. This modification utilises biotinylated PCR primers, which generate biotinylated linkers at an early stage in the SAGE protocol, thus allowing removal of the unwanted linkers by binding to streptavidin-coated magnetic beads at a later stage. The application of this modification resulted in the rapid generation of high ditag yields and clones with large average insert sizes.