In this experiment, the QIAGEN GeneRead DNAseq V2 Human Comprehensive Cancer Panel (cat. no. 181901) was used to generate PCR products [A] from a reference DNA sample. [B] A library was prepared from 25 ng of purified PCR product using the QIAseq 1-Step Amplicon Library Kit. Only 4 amplification cycles after library preparation were applied. Note that libraries display an appropriate size-shift and are free from adapter-adapter products (~125 bp).
After DNA extraction and purification with an appropriate kit, target enrichment is performed with the QIAGEN Multiplex PCR Kit, GeneRead DNAseq Targeted Panels V2 or other gene panels and PCR products. NGS library construction is performed with the QIAseq 1-Step Amplicon Library Kit and data is analyzed with the CLC Genomics or Biomedical Workbench.
The QIAseq 1-Step Amplicon Library Kit offers substantial time savings over standard targeted resequencing library
preparations. Purified amplicons from multiplexed PCR or gene panels are converted to sequencing libraries by employing a 30-minute, one-tube library construction step. The libraries are purified with an easy and automatable size selection protocol, and the entire procedure can be performed at room temperature. For low-input applications, libraries can be amplified using the included HiFi PCR Master Mix.