When it comes to achieving a clean template for your Sanger sequencing reaction, without losing precious samples, our sequencing ExoSAP-IT PCR product cleanup reagent delivers time and time again. Whether you’re looking for faster turnaround times or high-throughput, you can rely on our DNA sequencing PCR cleanup regaents to deliver the data quality and sequencing accuracy you need to move forward with complete peace of mind.
Before the PCR product is ready for Sanger sequencing, you must undertake a PCR cleanup of your reaction mixtures to prevent unincorporated primers and dNTPs from interfering with your results. If you overlook this vital step, you risk compromising the Sanger sequencing process and your abililty to effectively read DNA sequence base by base.
By offering a straightforward protocol and 100% recovery of both short and long amplicons, ExoSAP-IT PCR product cleanup reagent serves as a trusted tool in the researcher’s kit, allowing finite samples to be sufficiently conserved and productivity and process to be fine-tuned as part of a single, end-to-end workflow.
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.
Accelerate result accuracy with confidence with ExoSAP-IT Express reagent. Our one-step solution that delivers higher yields and full PCR product recovery.
Applied Biosystems ExoSAP-IT Express reagents are proprietary mixtures of exonuclease I combined with shrimp alkaline phosphatase (SAP) in a specially formulated buffer. Informed by decades of results, these reagents expertly remove excess primers and dNTPs following PCR. Exonuclease I removes residual single-stranded primers and any extraneous single-stranded DNA produced during PCR, while SAP removes the remaining dNTPs from the PCR mixture that may inhibit 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.
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.
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 method removes excess primers and dNTPs and does not interfere with downstream applications. No matter the parmeters of your project, our suite of PCR cleanup solutions deliver the flexibility and agility you need to optimize your workflow with confidence. Use our selection guide to determine which formulation is best for your next experiment.
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)|
|Freezes at |
|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|
Learn more about the superior workflow advantages of enzymatic purification technology.
ExoSAP-IT and ExoSAP-IT Express PCR cleanup reagents offer significant workflow advantages compared to magnetic beads or spin columns, while achieving similar results.
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.