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.