A comparison of two methods of template amplification for next generation sequencing: Implications of polyclonal formation on DNA sequence for several cancer tissues

<p>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.</p>

Name thesis:degree_name

Master of Science (MS)

Level thesis:degree_level

Open Access Thesis

Discipline thesis:degree_discipline

Health Sciences

Year dc:date.available

2016

Author dc:creator

• Aldilaimi, Akram

Contributors dc:contributor

• Irwin Martin
• David Kass
• Joseph Paulauskis

• DNA Sequence
• Genomic sequencing
• Next Generation Sequencing
• PCR duplications
• Personalized Medicine
• Polyclonal formation
• Medicine and Health Sciences

Repository record dc:identifier

https://commons.emich.edu/theses/790

OAI identifier oai:identifier

oai:commons.emich.edu:theses-2168