Applied Biosystems™ HT ExoSAP-IT™ Fast High-Throughput PCR Product Cleanup

Like the original formulation, HT ExoSAP-IT Fast reagent is a mixture of Exonuclease I and Shrimp Alkaline Phosphatase for the enzymatic removal of excess primers and dNTPs following a PCR reaction. This enzymatic cleanup reagent has a decreased viscosity yet maintains the same convenience and stability that you have come to expect from the ExoSAP-IT product line. As with all products in the ExoSAP-IT family, HT ExoSAP-IT Fast reagent allows PCR cleanup with a single pipetting step.

HT ExoSAP-IT Fast reagent is scalable for PCR cleanup from a single tube up to a 384-well plate PCR reaction.

• 14 minute protocol is half the time of standard protocols
• One-tube PCR cleanup — add HT ExoSAP-IT Fast reagent directly to PCR product
• 100% sample recovery — no loss of PCR products regardless of the fragment size
• Stable at 4°C for one month
• Removes excess primers and dNTPs — does not interfere with downstream applications
  • sequencing
  • SNP analysis
  • single base extension
  • fragment analysis

Adding to our family of patented ExoSAP-IT products, HT ExoSAP-IT Fast High-Throughput PCR Cleanup reagent quickly provides accurate and consistent results in high-throughput applications. HT ExoSAP-IT Fast reagent is over 50% faster than standard HT ExoSAP-IT reagent with a 14 minute total protocol time.

• Add it directly to the PCR product and incubate at 37°C for just 7 minutes.
• After PCR treatment, inactivate it by simply heating to 80°C for another 7 minutes.

This product is ideal for high volume labs that require quick sample turn around. With such a brief and simple protocol, proper sample cleanup never needs to be sacrificed due to processing or instrument time constraints. Ensure the highest quality Sanger sequencing results while keeping sample processing moving quickly and efficiently.

PCR products cleaned up with HT ExoSAP-IT Fast reagent prior to automated sequencing displayed superior sequencing results compared to the untreated equivalent (Fig. 2).

HT ExoSAP-IT Fast reagent displayed excellent stability in all formats tested including vials, 8-tube strips, and 96-well plates. The product showed no loss in function after 10 freeze thaw cycles (Fig. 3), and is stable at variable temperatures for extended periods of time compared to enzyme mix equivalents. With excellent stability and a greater than 50% faster protocol, HT ExoSAP-IT Fast reagent is ideal for high-throughput assays where robotic platforms are utilized and large volumes of samples are processed in successive testing runs.

Collectively, these results clearly demonstrate the effectiveness of HT ExoSAP-IT Fast reagent to ensure the highest quality sequences while keeping sample processing moving quickly and efficiently.