Simple: single-step PCR Product Clean-Up with USB ExoSAP-IT reagent
- Accuracy — achieve high data quality and sequencing accuracy, even with long read lengths
- 100% sample recovery — enzymatic PCR cleanup protocol, no loss of PCR products, regardless of the fragment size
- Convenience and speed — one-tube/one-step PCR cleanup
- Green option — generate less waste with our single-tube solution to PCR purification vs. columns
- Eliminate spin columns — decrease time and expense while increasing yield
- Cost savings — save as much as 50% vs. some spin column methods
- Downstream applications — DNA sequencing, TA cloning, and SNP analysis
- The gold standard in enzymatic PCR cleanup — ExoSAP-IT reagent has been referenced in thousands of publications since 2000; the market leader in enzymatic PCR clean-up with an established reputation for consistency and quality
ExoSAP-IT reagent is designed for simple, quick PCR clean-up for downstream applications, such as DNA sequencing or Single Nucleotide Polymorphism (SNP) analysis. When PCR amplification is complete, any unconsumed dNTPs and primers remaining in the PCR product mixture will interfere with these methods. ExoSAP-IT reagent removes these contaminants.
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How it works
ExoSAP-IT reagent is added directly to the PCR product and incubated at 37°C for 15 minutes. After PCR treatment, ExoSAP-IT reagent is inactivated simply by heating to 80°C for 15 minutes.
ExoSAP-IT single-step PCR clean-up utilizes two hydrolytic enzymes, Exonuclease I and Shrimp Alkaline Phosphatase, together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exo I removes residual single-stranded primers and any extraneous single-stranded DNA produced in the PCR. SAP removes the remaining dNTPs from the PCR mixture.
Achieve high data quality from PCR products
ExoSAP-IT reagent may be used as an effective clean-up method prior to fluorescent or radioactive DNA sequencing, SNP analysis, or any other application requiring a PCR product free of excess nucleotides and primers.
The data below shows the results of downstream processing with PCR product cleaned up by ExoSAP-IT reagent, an alternative enzymatic reagent, or a spin column method. The ExoSAP-IT reagent data shows superior results with no miscalls or frame shift mutations.
Fig. 1. Fluorescent sequencing results of a 1,007 bp PCR product. A 1,007 bp fragment was amplified and treated with ExoSAP-IT reagent (above) or an alternate enzymatic reagent (below) and sequenced. Pherograms revealed no miscalls with ExoSAP-IT reagent but four miscalls at position 25, 26, 30, and 39 with the alternate enzymatic reagent. 5 µl of PCR product was treated with 2 µl ExoSAP-IT reagent or the alternate enzymatic method per its protocol. Note: Two of the four miscalls are frame-shifts.
Fig. 2. Fluorescent sequencing results of a 100 bp pUC18 PCR fragment sequenced with a -20 Fwd primer using fluorescent sequencing reagents. PCR clean-up performed with: (a) ExoSAP-IT reagent; (b) a column designed for PCR clean-up. Base miscalls in (b) are due to inherently low yields of short PCR products when using columns.
Use the ExoSAP-IT method for PCR Clean-Up and see the difference 100% can make.