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ExoSAP-IT™ from GE Healthcare is a very simple PCR purification method that utilizes two hydrolytic enzymes, Exonuclease I and Shrimp Alkaline Phosphatase to remove any unconsumed dNTPs and primers prior to cycle sequencing. The Exonuclease I degrades residual single-stranded primers and any extraneous single-stranded DNA produced by the PCR. The Shrimp Alkaline Phosphatase hydrolyzes remaining dNTPs from the PCR mixture that would interfere with the sequencing reaction.

ExoSAP-IT™ (1 microlitre) is added directly to unpurified PCR product (5 microlitres). The enzymes are activated at 37°C for 30 minutes and then inactivated simply by heating to 80°C for 15 minutes. ExoSAP-IT consists of one pipetting step and two incubations (using a thermal cycler) and the reagent costs approximately $1.00 per sample (based on µL/reaction usage).

ExoSAP-IT™ company protocol

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