ExoSAP-IT Express PCR Cleanup Reagents
- Five-minute protocol—fast turnaround time
- One-tube, one-step PCR cleanup— add reagent directly to PCR product
- Novel enzyme technology— irreversibly inactivated in just one minute at 80°C
- Complete sample recovery—100% recovery of PCR products, regardless of amplicon length
- Eliminates 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 for manual or automated workflows
- Optional tracking dye—never lose track of reagent addition during pipetting
ExoSAP-IT Express reagents are patented mixtures of exonuclease I combined with shrimp alkaline phosphatase (SAP) in a specially formulated buffer, which remove excess primers and dNTPs following PCR. Exonuclease I removes residual single-stranded primers and any extraneous single-stranded DNA produced during PCR. SAP removes the remaining dNTPs from the PCR mixture that may interfere with subsequent reactions. Adding ExoSAP-IT Express reagent directly to the PCR product eliminates transfer steps to tubes, wells, or columns and conserves amplicons, helping to reduce the chance of cross-contamination; no further processing is needed. After enzymatic cleanup with ExoSAP-IT Express reagent, primers and dNTPs will no longer interfere with DNA sequencing or other downstream applications.
ExoSAP-IT Express: fastest PCR cleanup method
ExoSAP-IT Express reagent offers rapid turnaround times and improved efficiency of resource use while delivering the same superior cleanup as the original ExoSAP-IT reagent. Eliminate spin columns, magnetic beads, sedimentation, filtration, and gel purifications—the novel ExoSAP-IT technology allows for a significant reduction in sample cleanup time with minimal steps, providing the simplest workflow (Figure 1).
Treat 5 μl of PCR product with 2 μl of ExoSAP-IT Express reagent. Treatment is carried out at 37°C for 4 minutes followed by an incubation period at 80°C for 1 minute to irreversibly inactivate both enzymes. Once enzyme inactivation is complete, your PCR products are ready for downstream applications such as sequencing (Sanger/NGS), fragment analysis, SNP analysis, in vitro transcription, or single base extension (Figure 2).
Figure 2. Enzymatic cleanup of PCR reaction with ExoSAP-IT Express reagent.
The original ExoSAP-IT formulation is considered the gold standard in enzymatic PCR cleanup. ExoSAP-IT PCR product cleanup has been cited in over 10,000 publications.
- 30 minutes cleanup protocol
- Recommended for processing 1-96 samples at a time
- Best cost per reaction
Applied Biosystems™ CleanSweep™ and ExoSAP-IT reagents are interchangeable in both function and performance. A direct comparison of sequencing data obtained after cleanup with ExoSAP-IT and CleanSweep reagents shows that both sets of data are equivalent in contiguous read length (CRL), quality score (QV), and peak under peak (PUP) (Figure 3).
Figure 3. Sequencing traces after enzymatic cleanup with (A) CleanSweep reagent, (B) ExoSAP-IT reagent, and (C) buffer only (negative control).
HT ExoSAP-IT™ High-Throughput PCR Product Cleanup is an alternative formulation of ExoSAP-IT reagent specifically designed for the unique requirements of high-throughput, automated platforms, and multi-channel pipettes. HT ExoSAP-IT reagent has both a longer lifetime at higher temperatures and a decreased viscosity for robotic pipetting.
Like ExoSAP-IT for PCR Product Cleanup (Cat. No. 78200), HT ExoSAP-IT reagent is a mixture of exonuclease I and shrimp alkaline phosphatase (SAP) which removes excess primers and dNTPs following a PCR reaction in a single incubation. HT ExoSAP-IT reagent possesses greater thermal stability and lower viscosity which provides greater ease-of-use in automated 96- and 384-well formats.
For Research Use Only. Not for use in diagnostic procedures.