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Applied Biosystems™ ExoSAP-IT™ Express PCR Product Cleanup
Description
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.
Specifications
Specifications
| Content And Storage | Shipped on dry ice. Store at -20°C. |
| Format | 8 Tube Strip, 96-well Plate, Tube |
| No. of Reactions | 20 Reactions |
| Quantity | 40 μL |
| Test Time | 5 min. |
Frequently Asked Questions (FAQs)
The HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup reagent is designed for automated handlers as it has a lower viscosity than standard ExoSAP-IT PCR Product Cleanup reagent, which contains some glycerol.
The Quant-iT PicoGreen dsDNA Assay Kit is recommended for quantifying PCR products that have been cleaned with ExoSAP-IT reagent. ExoSAP-IT PCR cleanup results in 100% recovery regardless of amplicon length, and if the PCR product was quantified prior to ExoSAP-IT cleanup, additional quantification is not necessary. We do not recommend using a spectrophotometer as degraded products will be included in the quantification.
For use with reverse transcription, ExoSAP-IT does not degrade the DNA-RNA hybrid. However, it will degrade single-stranded cDNA if a 3' -OH end is present.
The tracking dyes used in the DreamTaq Hot Start Green PCR Master Mix have not been shown to interfere with functionality of the ExoSAP-IT cleanup reagents.
The proofreading function of HiFi DNA polymerases can cause partial degradation of PCR products during extended incubation at 80°C. ExoSAP-IT Express reagent is recommended for HiFi PCR cleanup because it only requires 1 minute at 80°C for inactivation.
Safety and Handling
For Research Use Only. Not for use in diagnostic procedures.
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