PCR Cleanups

PureIT ExoZAP PCR CleanUp improves quality of DNA sequencing

Amplification of a 866 bp sized PCR product was performed using TEMPase DNA Polymerase in Ammonium Buffer. The PCR products was spiked with primers and dNTP’s followed by treatment (A) without PureIT ExoZAP PCR CleanUp (B) with PureIT ExoZAP PCR CleanUp (2 + 3 min) (C) with another enzymatic PCR clean-up reagent from a competitor (1 + 4 min). The samples were analyzed in triplicates and according to manufactures recommendations. The results of Sanger sequencing between de samples was compared by plotting the number of base calls in graphs showing a quality value score (Phred score). The PCR products that were treated with PureIT ExoZAP resulted in a significantly improved quality of DNA sequencing. Next to this, PureIT ExoZAP PCR CleanUp showed comparable PCR clean-up results to the competitor’s clean-up. *Data with Quality Values below 10 are not shown. 

The Sanger sequencing results of the 866 bp sized PCR product are displayed above. The upper electropherogram is after treatment with PureIT ExoZAP PCR CleanUp and the lower electropherogram is untreated. The displayed region is from 250 – 300 bases of the sequence. Treating the spiked PCR product with PureIT ExoZAP PCR CleanUp resulted in a significantly improved Sanger sequencing quality. Signal intensities are enhanced and background interference and base miscalls are eliminated.