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Nucleic Acids Research, Vol 26, Issue 14 3445-3446, Copyright © 1998 by Oxford University Press

ARTICLES

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

J Powell
The Richard Dimbleby Department of Cancer Research, I.C.R.F. Laboratory, Rayne Institute, 4th Floor Lambeth Wing, St Thomas's Hospital, Lambeth Palace Road, London SE1 7EH, UK. powell@icrf.icnet.uk

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.
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