HT ExoSAP-IT™ High-Throughput PCR Product Cleanup

HT ExoSAP-IT High-Throughput PCR Product Cleanup is an alternative formulation of ExoSAP-IT reagent specifically designed for the unique requirements of high-throughput, automated platforms, and multi-channel pipettes. HT ExoSAP-IT reagent has both a longer lifetime at higher temperatures and a decreased viscosity for robotic pipetting.

Like ExoSAP-IT for PCR Product Cleanup (PN 78200), HT ExoSAP-IT reagent is a mixture of Exonuclease I and Shrimp Alkaline Phosphatase (SAP) which removes excess primers and dNTPs following a PCR reaction in a single incubation. HT ExoSAP-IT reagent possesses greater thermal stability and lower viscosity which provides greater ease-of-use in automated 96- and 384-well formats.

• One-tube PCR cleanup — Add HT ExoSAP-IT reagent directly to PCR product
• Exceptional accuracy — Achieve high quality data even with long read lengths
• 10% sample recovery — No loss of PCR products regardless of the fragment size
• Simple, single-step — Replaces the multiple steps and wasted time inherent with beads or columns
• High-throughput processing — Even faster time to results with a low viscosity formulation, allowing robotic pipetting
• Removes excess primers and dNTPs — Does not interfere with downstream applications
  • sequencing
  • SNP analysis
  • single base extension
  • fragment analysis
  • in vitro transcription
• Scalable — Treat reaction volumes from 5 μL to 5 L
• Convenient packaging — Available in 8-tube strips and 12 strips in a 96-well plate
• Highly stable — Retains full functional activity at 4°C for one week and at 25°C for 8 hours

HT ExoSAP-IT reagent is added directly to the PCR product and incubated at 37°C for 15 minutes. After PCR treatment, it is inactivated simply by heating to 80°C for 15 minutes. Our high-throughput formulation is designed for robotic pipetting and multi-channel pipettes. HT ExoSAP-IT reagent has a lower viscosity than the standard mix and is available in an 8-tube strip format for ease of use.

HT ExoSAP-IT reagent utilizes two hydrolytic enzymes, Exonuclease I and 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.

Simple: Single-step

HT ExoSAP-IT reagent requires only one pipetting step and two incubations. Just add HT ExoSAP-IT reagent to the PCR product and within 30 minutes sequencing, SNP analysis, single base extension, or fragment analysis can be performed. Enzymatic removal of excess primers and unincorporated nucleotides occurs in one easy step by using HT ExoSAP-IT reagent in a single tube, 8-tube strip, or 12 strips in a 96-well low skirted PCR plate. Our new formatting makes this an ideal product for use with robotics.

Exceptional accuracy with HT ExoSAP-IT

Achieve high data quality and sequencing accuracy with HT ExoSAP-IT, even with long read lengths. Sequence reads of PCR products treated with HT ExoSAP-IT were on average 50+ bases longer than samples treated with competitor products. Using human genomic DNA, a 1007 bp fragment was sequenced with no miscalls. The size limitations associated with alternative PCR cleanup methods are not a factor with HT ExoSAP-IT. Phred 20 values over the entire length of a 970 bp sequence was 822 ± 9 with HT ExoSAP-IT-treated samples and only 776 ± 83 with samples treated with Agencourt™ AMPure™ XP.

No sample loss

Use of HT ExoSAP-IT eliminates all gel or column purifications, sedimentations, filtrations, beads, and/or magnetic separations⁽¹⁾. There is 100% recovery of both short and long PCR products with HT ExoSAP-IT.

HT ExoSAP-IT-treated PCR product is stable

Ht ExoSAP-IT reagent is rigorously tested and subject to strict quality control. No product degradation occurred after storage of HT ExoSAP-IT treated PCR product for one week at 24°C.

References:

1. DUGAN, K. A., LAWRENCE, H. S., HARES, D. R., FISHER, C. L. AND BUDOWLE B. (2002) J. Forensic Sci 47, 811-818.
2. HANKE, M. AND WINK, M. (1994) BioTechniques 17, 858-860.
3. MU, J., DUAN, J., MAKOVA, K., JOY, D., HUYNH, C., BRANCH, O., LI, W. AND SU, X. (2002) Nature 418, 323-326.
4. SILVA, JR., W. A., COSTA, M. C. R., VALENTE, V., DE FREITAS SOUSA, J., PACÓ-LARSON, M. L., ESPREAFICO, E. M., CAMARGO, S. S., MONTEIRO, E., DE JESUS, A., HOLANDA, M. A., ZAGO, M. A., SIMPSON, A. J. G. AND NETO, E. D. (2001) BioTechniques 30, 537-542.
5. WERLE, E., SCNEIDER C., RENNER, M., VÖLKER, M. AND FIEHN, W. (1994) Nucleic Acids Res. 22, 4354-4355.