ExoSAP-IT PCR Product Cleanup Reagent | Thermo Fisher Scientific

High-quality, accurate results, faster turnaround time

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.

Why is a clean template for your Sanger sequencing reaction important?

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.

Learn more: watch our Seq it Out video

Brochure: ExoSAP-IT PCR cleanup reagents

Learn more about how our ExoSAP-IT PCR Product Cleanup Reagent ensures quality sequencing results in a fraction of the time.

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How does ExoSAP-IT PCR product cleanup reagent optimize workflow and preserve sample integrity?

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.

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.

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ExoSAP-IT Express—one tube, one-step

Accelerate result accuracy with confidence with ExoSAP-IT Express reagent. Our one-step solution that delivers 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
Consolidated ordering from one trusted partner

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Overview

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.

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.

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An accelerated, optimized workflow you can rely on

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 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
Format	Single tube 8-tube strips	Single tube	Single tube 8-tube strips 96-well plate
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 –20°C	No	No	Yes
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
	Purchase	Purchase	Purchase

Comparison to alternative PCR cleanup methods

Learn more about the superior workflow advantages of enzymatic purification technology.

ExoSAP_IT_Express_PCR_Product_Cleanup_Comparison

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.

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