ExoSAP-IT Express PCR Product Cleanup with Tracking Dye
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Applied Biosystems™

ExoSAP-IT Express PCR Product Cleanup with Tracking Dye

Our fastest PCR cleanup method - just 5 minutes to superior sequencing results. Now available with optional tracking dye.ExoSAP-IT™ ExpressMás información
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Número de catálogoN.º de reacciones
A35004500 reacciones
A350051920 reacciones
Número de catálogo A35004
Precio (CLP)
608.290
Each
Añadir al carro de la compra
N.º de reacciones:
500 reacciones
Precio (CLP)
608.290
Each
Añadir al carro de la compra

Our fastest PCR cleanup method - just 5 minutes to superior sequencing results. Now available with optional tracking dye.

ExoSAP-IT™ Express reagent:
• 5 minute protocol—Fastest turnaround time to results
• One-tube, one-step PCR cleanup—Add ExoSAP-IT Express directly to PCR product
• Novel enzyme technology—Irreversibly inactivated in just one minute at 80°C
• Conserve PCR samples—100% recovery of PCR products, regardless of amplicon length
• Eliminate spin columns or beads—Significantly decreases 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 from manual pipetting to automated workflows
• Go green—Generate less waste with a single-tube solution to PCR cleanup
• Optional tracking dye—Never lose track of reagent addition during pipetting

Overview
It is necessary to remove excess primers and unincorporated nucleotides from the PCR reaction prior to sequencing. An enzymatic cleanup method offers precision and speed over technologies such as spin columns or beads. ExoSAP-IT Express reagent consists of two hydrolytic enzymes, a modified Exonuclease I and Shrimp Alkaline Phosphatase (SAP), in a specially formulated buffer. It is used to remove excess primers and dNTPs from a PCR mixture with no interference in downstream applications. This unique, highly stable, one tube solution allows for greater confidence and superior sequencing results (Fig. 1).

Innovative solution
Affymetrix scientists engineered Exonuclease I to have increased heat lability, allowing for inactivation in only 1 minute (Fig. 2). ExoSAP-IT Express reagent is added directly to the PCR product for a 4 minute cleanup at 37°C, followed by a 1 minute inactivation at 80°C, which provides accurate and consistent results in just 5 minutes.

Fastest PCR cleanup method
This novel technology allows for a significant reduction in sample cleanup time with minimal steps, providing the simplest workflow (Fig. 3). ExoSAP-IT Express reagent offers rapid turnaround times and improved efficiency of resources while delivering the quality that scientists have become accustomed to when using ExoSAP-IT for PCR product cleanup.

Optional tracking dye
ExoSAP-IT Express reagent is also available premixed with a tracking dye for added convenience. This inert, blue color dye
offers visual confirmation that the reagent has been added to samples for PCR cleanup during the pipetting process. The dye does not interfere with downstream applications such as subsequent PCR and sequencing.

Conserve PCR samples — simple one step, 100% recovery
The ExoSAP-IT Express enzymatic cleanup method is designed to minimize errors by reducing your protocol to a single pipette step, allowing for automated or manual processing from a single tube or microtiter well. ExoSAP-IT Express reagent outperforms the competition by ensuring 100% recovery of all amplicon sizes, both short and long (Table 1).

ExoSAP-IT Express PCR Product Cleanup is active in commonly used PCR buffers, so no buffer exchange is required. After treatment, ExoSAP-IT Express reagent is inactivated by heating at 80°C for 1 minute. The treated PCR products are then ready for subsequent analysis in applications that require DNA to be free of excess primers and nucleotides.

Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones
Para utilizar con (aplicación)Purificación de PCR
N.º de reacciones500 reacciones
Línea de productosExoSAP-IT
Tipo de productoPCR Product Cleanup
Forma de proteínaRecombinante
Cantidad500 reactions
Categoría de investigaciónEnfermedades genéticas
Condiciones de envíoHielo seco
Unit SizeEach
Contenido y almacenamiento
Se envía con hielo seco. Almacenar a -20 °C.

Preguntas frecuentes

Can ExoSAP-IT PCR cleanup reagents be used with automatic handling systems?

The HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup reagent is designed for automated handlers as it has a lower viscosity than standard ExoSAP-IT PCR Product Cleanup reagent, which contains some glycerol.

After purification with ExoSAP-IT PCR cleanup reagents, what is the recommended quantification method?

The Quant-iT PicoGreen dsDNA Assay Kit is recommended for quantifying PCR products that have been cleaned with ExoSAP-IT reagent. ExoSAP-IT PCR cleanup results in 100% recovery regardless of amplicon length, and if the PCR product was quantified prior to ExoSAP-IT cleanup, additional quantification is not necessary. We do not recommend using a spectrophotometer as degraded products will be included in the quantification.

Can ExoSAP-IT PCR cleanup reagents be used for removal of primers after first-strand cDNA synthesis?

For use with reverse transcription, ExoSAP-IT does not degrade the DNA-RNA hybrid. However, it will degrade single-stranded cDNA if a 3' -OH end is present.

Will the two tracking dyes used for visualization in the DreamTaq Hot Start Green PCR Master Mix interfere with the ExoSAP-IT PCR cleanup reagents?

The tracking dyes used in the DreamTaq Hot Start Green PCR Master Mix have not been shown to interfere with functionality of the ExoSAP-IT cleanup reagents.

Can ExoSAP-IT PCR cleanup reagents be used to cleanup PCR products amplified using High Fidelity (HiFi) DNA polymerases?

The proofreading function of HiFi DNA polymerases can cause partial degradation of PCR products during extended incubation at 80°C. ExoSAP-IT Express reagent is recommended for HiFi PCR cleanup because it only requires 1 minute at 80°C for inactivation.