ExoSAP-IT™ PCR Product Cleanup

Features and Benefits of the ExoSAP-IT™ PCR Product Cleanup
• Conserves PCR samples — 100% recovery of both short and long PCR products
• One tube/one step PCR cleanup — Add ExoSAP-IT reagent directly to PCR product
• Eliminates spin columns — Decreases time and expense while increasing yield
• Removes contaminating primers and dNTPs — No interference in downstream applications
• Scalable — Economical for high-throughput purification
• Simple processing — Robotic-friendly; Replaces beads, filtrations, and plates
• Generates less waste than columns

ExoSAP-IT reagent is designed for simple, quick PCR cleanup 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 removes these contaminants.

ExoSAP-IT is added directly to the PCR product and incubated at 37°C for 15 minutes (Fig. 1). After PCR treatment, ExoSAP-IT is inactivated simply by heating to 80°C for 15 minutes.

ExoSAP-IT single-step PCR cleanup utilizes two hydrolytic enzymes, Exonuclease I and Shrimp Alkaline Phosphatase (SAP), together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exonuclease 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.

Rapid PCR Product Cleanup Protocol
ExoSAP-IT requires only one pipetting step and two incubations. Just add ExoSAP-IT to the PCR product and within 30 minutes sequencing or SNP analysis can be performed.

Simple: Single-Step
The method is designed to require a minimum of ‘hands-on’ time. Enzymatic removal of excess primers and unincorporated nucleotides occurs in one easy step by using ExoSAP-IT reagent in a single tube or microtiter well. Only simple pipette transfers are required, therefore, many samples can be processed at once, either manually or with robotics.

No Sample Loss
Use of ExoSAP-IT reagent eliminates all gel or column purifications, sedimentations, filtrations, beads, and/or magnetic separations. There is 100% recovery of both short and long PCR products with ExoSAP-IT (Fig. 2).

Achieve High Data Quality from PCR Products
ExoSAP-IT reagent may be used as an effective cleanup method prior to fluorescent or radioactive DNA sequencing, SNP analysis, or any other application requiring a PCR product free of excess nucleotides and primers.

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