Preparing Polymerase Chain Reaction (PCR) Products for Capillary Sequencing

Abstract

This protocol describes the preparation of amplified products for use in Sanger-based capillary DNA sequencing, for example, to verify a clone or a construct. Amplified samples, subsequently treated with a mixture of exonuclease and shrimp alkaline phosphatase to remove unincorporated primers and dNTPs left from polymerase chain reaction (PCR), may be used directly for sequencing. This protocol relies on the use of the Biomek FX Workstation or a multichannel pipettor.