Project

qPCR system for NGS library quality control, gene expression measurement, hybridization curve, and melting curve analysis

Code
BOF/BAS/2022/008
Duration
15 December 2022 → 14 December 2024
Funding
Regional and community funding: Special Research Fund
Research disciplines
  • Natural sciences
    • Animal genetics
    • Animal immunology
  • Medical and health sciences
    • Immunogenetics
    • Clinical genetics and molecular diagnostics
    • Microbiome
    • Immunogenetics
    • Microbiome
    • Developmental biology
    • Epigenetics
    • Genetics
    • Transcription and translation
    • Clinical genetics and molecular diagnostics
    • Biomarker discovery
    • Immunogenetics
    • Clinical genetics and molecular diagnostics
    • Microbiome
    • Epigenetics
    • Genetics
    • Transcription and translation
  • Agricultural and food sciences
    • Veterinary embryology
    • Veterinary genetics
    • Veterinary microbiology
    • Veterinary oncology
    • Veterinary reproduction and obstetrics
Keywords
hybridisation curve analysis melt curve analysis gene expression analysis next generation sequencing qPCR
 
Project description

In the UGent NXTGNT NGS sequencing laboratory, the Roche LightCycler 480 II qPCR system has served dozens of UGent researchers across several faculties, contributing to hundreds of publications. Also, dozens of external universities, institutions, and commercial partners have benefitted from the system. Most of these researchers indirectly used the system by performing NGS library preparation in the NXTGNT lab. The qPCR system performs quality control and quantifies sequenceable fragments as a step in every NGS library preparation. Additionally, the system is used by several groups to perform gene expression analysis, CRISPR-Cas9 cleavage efficiency, and high resolution melting curve analysis.

The existing system has been in use almost daily since 2009 and needs to be replaced. Due to wear and tear, the instrument often shows hardware defects, resulting in unreliable measurements. The frequent and costly repairs cannot be justified anymore due to the system's age.