Open Access Thesis
Master of Science (MS)
Next-Generation Sequencing (NGS) technology has advanced the field of personalized medicine by predicting effective treatments for cancer patients using genomic sequence data, including the detection of oncogenes (genes involved in cancer development). Simultaneous amplification of multiple DNA templates (referred to as polyclonal formation) is a primary disadvantage when preparing templates for NGS resulting in redundancies in DNA sequences or nonspecific noise in the sequencing. This study was conducted to compare DNA templates for sequencing prepared using two instruments the Thermo Fisher Ion Chef (IC) and Thermo Fisher OneTouch-2 (OT2). Six sequencing metrics obtained from 114 sequencing trials for evaluation: polyclonal reads, total sequencing reads, empty microcell well reads, no-template reads, useable number reads, and library number reads. A comparison of mean sequencing metrics between the IC and OT2 methods established that for four of the six metrics, IC was the preferred operation for DNA template preparation due to the less polyclonal formation.
Aldilaimi, Akram, "A comparison of two methods of template amplification for next generation sequencing: Implications of polyclonal formation on DNA sequence for several cancer tissues" (2016). Master's Theses and Doctoral Dissertations. 790.