ExoSAP-IT™ Express PCR Product Cleanup, 40 μL - FAQs

View additional product information for ExoSAP-IT™ Express PCR Product Cleanup - FAQs (75001.1.ML, 75001.1.EA, 75001.40.UL, 75001.4X.1.ML, 75001.200.UL, 75001.10.ML)

8 product FAQs found

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.

Can ExoSAP-IT-treated PCR products be used in cloning applications?

ExoSAP-IT-treated PCR products can be used in TA cloning, as the exonuclease I in ExoSAP-IT reagent will not degrade the single A-overhang required for TA cloning.

How do the ExoSAP-IT PCR cleanup reagents help improve sequencing results?

ExoSAP-IT treatment prior to sequencing helps eliminate miscalls, improves quality scores, increases continuous read length (CRL), and decreases background interference. This yields sequencing results with greater confidence, consistency, and accuracy.

What are the differences between the ExoSAP-IT PCR cleanup reagents?

All of our ExoSAP-IT reagents have a similar function for PCR product cleanup, but differ in the areas listed below:

ExoSAP-IT Express PCR Product Cleanup
• Protocol time: 5 minutes
• Format: Single tube*, 8-tube strips, 96-well plate*
• Throughput level: Low to high; Recommended for processing any sample size
• Platform: Single- or multi-channel pipette, automated liquid handling platforms
• Freezes at -20°C: No
• Stability: -20°C for up to 2 years

ExoSAP-IT PCR Proudct Cleanup (our original formulation)
• Protocol time: 30 minutes
• Format: Single tube
• Throughput level: Low to mid; Recommended for processing 1-96 samples at a time
• Platform: Single-channel pipette
• Freezes at -20°C: No
• Stability: -20°C for up to 2 years

HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup (for automated liquid handlers)
• Protocol time: 14 minutes
• Format: Single tube, 8-tube strips, 96-well plate
• Throughput level: High; Recommended for processing ≥96 samples at a time
• Platform: Automated liquid handling platforms
• Freezes at -20°C: Yes
• Stability: -20°C for up to 2 years; once thawed, stable at 4°C for 1 month and RT for 2 days

*Optional tracking dye available (Cat. No. 75002)