High-quality, accurate results, faster turnaround time
It is important to achieve a clean template for your Sanger sequencing reaction without losing precious samples. Our sequencing Exo-SAP-IT PCR product cleanup reagent provides fast, easy, and convenient solutions for your PCR cleanup needs. Whether you require the fastest turnaround time or high-throughout, our DNA sequencing PCR cleanup reagents ensure data quality and sequencing accuracy.
Before the PCR product is ready for Sanger sequencing, you must do a PCR cleanup step. Because Sanger sequencing is a highly accurate technique for you to read DNA sequence base by base, it is very important to clean up your reaction mixtures so that those unincorporated primers and dNTPs won’t interfere with your results.
Learn more: watch our Seq it Out video
With its simple protocol and 100% recovery of both short and long amplicons, ExoSAP-IT PCR product reagent enables researchers to conserve limited samples and improve workflow efficiency.
Application note: Use of ExoSAP-IT PCR Product Cleanup Reagent in NGS
While Sanger sequencing–based methods remain popular for validation and long contiguous DNA sequence reads (>500 nucleotides), many genomic analysis applications are transitioning to next-generation sequencing (NGS) technology for its scalability and affordability for sequencing a large number of targets.
ExoSAP-IT Express—one tube, one-step
Achieve superior results with ExoSAP-IT Express reagent. Improve accuracy with higher yields and full PCR product recovery.
- 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—helps to significantly decrease 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—helps to avoid losing track of reagent addition during pipetting
Applied Biosystems ExoSAP-IT Express reagents are proprietary 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.
Figure 1. Enzymatic cleanup of PCR reaction with ExoSAP-IT Express reagent. 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.
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.
Product Spotlight: ExoSAP-IT Express PCR Cleanup Agent
In only five minutes, ExoSAP-IT Express PCR Cleanup Agent removes excess primers and unincorporated nucleotides from a PCR reaction. This enzymatic cleanup method offers precision, increased yield, and a faster turnaround time compared to alternative purification methods, such as spin columns or beads.
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 (see figure).
Figure 2. ExoSAP-IT Express reagent vs. other common methods.
Complete ExoSAP-IT PCR product cleanup portfolio—comprehensive solutions
ExoSAP-IT reagents offer a unique one-tube, one-step enzymatic method for PCR cleanup. All ExoSAP-IT reagents provide 100% recovery of PCR products regardless of the fragment sizes. This PCR cleanup method removes excess primers and dNTPs and does not interfere with downstream applications.
Whether you are looking for a fast turnaround time or high-throughput solution, we have a PCR cleanup reagent that will work for you. Use our selection guide to determine which formulation is best for your next experiment.
|ExoSAP-IT Express reagent||ExoSAP-IT reagent (original formulation)||HT ExoSAP-IT Fast High-Throughput reagent|
|Protocol time||5 minutes||30 minutes||14 minutes|
|Throughput level||Low to high; recommended for processing any sample size||Low to medium; recommended for processing 1–96 samples at a time||High; recommended for processing ≥96 samples at a time|
|Platform||Single- or multichannel pipette, automated liquid handling platforms (lower viscosity for better handling)||Single-channel pipette||Automated liquid handling platforms (lower viscosity for better handling)|
|Stability||–20°C for up to 2 years||–20°C for up to 2 years||–20°C for up to 2 years; once thawed, stable at 4°C for 1 month and at room temperature for 2 days|
Comparison to alternative PCR cleanup methods
Learn more about the superior workflow advantages of enzymatic purification technology.
Comparison of ExoSAP-IT and ExoSAP-IT Express reagents to alternative PCR cleanup methods
ExoSAP-IT and ExoSAP-IT Express PCR cleanup reagents offer significant workflow advantages compared to magnetic beads or spin columns, while achieving similar results.
Comparison of CleanSweep and ExoSAP-IT PCR clean-up agents
A side-by-side comparison of CleanSweep and ExoSAP-IT reagents that demonstrates ExoSAP-IT effectiveness and functional equivalency.
For Research Use Only. Not for use in diagnostic procedures.