Most next-generation sequencing (NGS) library construction workflows include one or more PCR amplification steps to (i) add functional elements (e.g., sample indices, molecular barcodes or flow cell oligo binding sites), (ii) enrich for sequencing-competent DNA fragments and/or (iii) generate a sufficient amount of library material for downstream processing. PCR is also applied for other purposes in NGS workflows. Whenever PCR is performed, care must be taken to select an appropriate enzyme and protocol parameters, to ensure high efficiency, minimize amplification bias and limit the introduction of sequence artifacts.
Roche Sequencing Solutions offers a family of KAPA HiFi Kits containing KAPA HiFi DNA Polymerase, engineered using directed evolution technology. The KAPA HiFi enzyme contributes to higher overall sequencing quality and economy by offering high efficiency, low bias amplification of DNA and cDNA for both NGS and PCR applications. This translates to more uniform coverage of difficult regions and lower duplication rates allowing researchers to unlock the potential of their samples. In addition, the KAPA HiFi family includes KAPA HiFi Uracil+ Kits, which contain a uracil-tolerant variant, KAPA HiFi Uracil+ DNA Polymerase. This enzyme still offers the same benefits enabled by KAPA HiFi, but is also tolerant to amplification of modified nucleic acids, including bisulfite-converted DNA.