Applied Biosystems™ ExoSAP-IT™ Express PCR Product Cleanup

Key features of ExoSAP-IT™ Express reagent:

• 5 minute protocol—Fastest turnaround time to results
• One-tube, one-step PCR cleanup—Add ExoSAP-IT Express directly to PCR product
• Novel enzyme technology—Irreversibly inactivated in just one minute at 80°C
• Conserve PCR samples—100% recovery of PCR products, regardless of amplicon length
• Eliminate spin columns or beads—Significantly decreases time and errors while increasing yield
• Compatible—No interference in downstream applications
• Scalable—Treat a wide range of volumes
• Multiple formats—Single tube, 8 tube strip, and 96-well plate formats offer flexibility from manual pipetting to automated workflows
• Go green—Generate less waste with a single-tube solution to PCR cleanup

Overview

It is necessary to remove excess primers and unincorporated nucleotides from the PCR reaction prior to sequencing.

An enzymatic cleanup method offers precision and speed over technologies such as spin columns or beads. ExoSAP-IT Express reagent consists of two hydrolytic enzymes, a modified Exonuclease I and Shrimp Alkaline Phosphatase (SAP), in a specially formulated buffer. It is used to remove excess primers and dNTPs from a PCR mixture with no interference in downstream applications. This unique, highly stable, one tube solution allows for greater confidence and superior sequencing results (Fig. 1).

Innovative solution

Our scientists engineered Exonuclease I to have increased heat lability, allowing for inactivation in only 1 minute (Fig. 2). ExoSAP-IT Express reagent is added directly to the PCR product for a 4 minute cleanup at 37°C, followed by a 1 minute inactivation at 80°C, which provides accurate and consistent results in just 5 minutes.

Fastest PCR cleanup method

This novel technology allows for a significant reduction in sample cleanup time with minimal steps, providing the simplest workflow (Fig. 3). ExoSAP-IT Express reagent offers rapid turnaround times and improved efficiency of resources while delivering the quality that scientists have become accustomed to when using ExoSAP-IT for PCR product cleanup.

Conserve PCR samples — simple one step, 100% recovery

The ExoSAP-IT Express enzymatic cleanup method is designed to minimize errors by reducing your protocol to a single pipette step, allowing for automated or manual processing from a single tube or microtiter well. ExoSAP-IT Express reagent outperforms the competition by ensuring 100% recovery of all amplicon sizes, both short and long (Table 1).

ExoSAP-IT Express PCR Product Cleanup is active in commonly used PCR buffers, so no buffer exchange is required. After treatment, ExoSAP-IT Express reagent is inactivated by heating at 80°C for 1 minute. The treated PCR products are then ready for subsequent analysis in applications that require DNA to be free of excess primers and nucleotides.