UNIT 15.2 Direct DNA Sequencing of PCR Products
Published Online: 1 NOV 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Dorit, R. L., Ohara, O., Hwang, C. B.-C., Kim, J. B. and Blackshaw, S. 2001. Direct DNA Sequencing of PCR Products. Current Protocols in Molecular Biology. 56:15.2:15.2.1–15.2.13.
- Published Online: 1 NOV 2001
- Published Print: OCT 2001
PCR products can be sequenced using either the dideoxy (Sanger) or chemical (Maxam-Gilbert) approaches. In the dideoxy methods presented here, the target sequence is amplified and an excess of one strand of the target sequence (relative to its complement) is then generated by “asymmetric PCR,” where one primer is present in vast excess over the other. This single-stranded product serves as the template for conventional dideoxy sequencing methods. Another procedure prepares PCR products for use as templates fes for characterizing unlabeled product by genomic sequencing and chemical sequencing of end-labeled products are also presented.