Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1999 Feb 1;27(3):917–918. doi: 10.1093/nar/27.3.917

Substantially enhanced cloning efficiency of SAGE (Serial Analysis of Gene Expression) by adding a heating step to the original protocol.

M Kenzelmann 1, K Mühlemann 1
PMCID: PMC148268  PMID: 9889294

Abstract

The efficiency of the original SAGE (Serial Analysis of Gene Expression) protocol was limited by a small average size of cloned concatemers. We describe a modification of the technique that overcomes this problem. Ligation of ditags yields concatemers of various sizes. Small concatemers may aggregate and migrate with large ones during gel electrophoresis. A heating step introduced before gel electrophoresis breaks such contaminating aggregates. This modification yields cloned concatemers with an average size of 67 tags as compared to 22 tags by the original protocol. It enhances the length of cloned concatemers substantially and reduces the costs of SAGE.

Full Text

The Full Text of this article is available as a PDF (34.7 KB).


Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES