Fabrication of DNA Microarrays by Digital Solid-Phase PCR in a Next Generation Sequencing Chip

Inhaltsverzeichnis (Table of Contents)

• Abstract
• 1. Introduction
• 2. State of the Art
• 3. Materials and Methods
• 3.1. DNA Microarray Fabrication
• 3.2. Immobilization of PCR Primers onto Microscope Slides
• 3.3. Solid-Phase PCR (SP-PCR) on Microscope Slides
• 3.4. On-Chip PCR
• 3.5. Digital Solid-Phase PCR (dSP-PCR) in a PicoTiterPlate (PTP)
• 3.6. DNA Microarray Replication
• 4. Results and Discussion
• 4.1. Immobilization of PCR Primers onto Microscope Slides
• 4.2. Solid-Phase PCR (SP-PCR) on Microscope Slides
• 4.3. On-Chip PCR
• 4.4. Digital Solid-Phase PCR (dSP-PCR) in a PicoTiterPlate (PTP)
• 4.5. DNA Microarray Replication
• 5. Conclusion
• 6. Outlook
• References

Zielsetzung und Themenschwerpunkte (Objectives and Key Themes)

This dissertation presents a novel process for the fabrication of DNA microarrays using a next-generation sequencing chip. The main objective is to develop a method for replicating DNA microarrays from a master microarray generated by digital solid-phase PCR in a PicoTiterPlate (PTP).

• Immobilization of PCR primers onto various materials
• Solid-phase PCR (SP-PCR) on different substrates
• Digital solid-phase PCR (dSP-PCR) in a PTP
• DNA microarray replication
• Minimizing carry-over DNA contamination in dSP-PCR

Zusammenfassung der Kapitel (Chapter Summaries)

• Chapter 1: Introduction This chapter introduces the concept of DNA microarrays and their applications. It discusses the limitations of traditional DNA microarray fabrication methods and highlights the potential of using next-generation sequencing chips for this purpose.
• Chapter 2: State of the Art This chapter provides a comprehensive overview of the existing technologies for DNA microarray fabrication, including the use of solid-phase PCR and digital PCR. It also reviews the different methods for immobilizing PCR primers onto surfaces.
• Chapter 3: Materials and Methods This chapter details the materials and methods used in the dissertation. It describes the fabrication process of DNA microarrays using a PicoTiterPlate (PTP) and the different steps involved, including primer immobilization, solid-phase PCR, and DNA microarray replication.
• Chapter 4: Results and Discussion This chapter presents the results of the experiments conducted in the dissertation. It analyzes the efficiency of different methods for primer immobilization, solid-phase PCR, and digital solid-phase PCR. It also discusses the challenges and solutions related to carry-over DNA contamination in dSP-PCR.

Schlüsselwörter (Keywords)

DNA microarrays, next-generation sequencing, solid-phase PCR, digital PCR, PicoTiterPlate (PTP), primer immobilization, DNA replication, carry-over DNA, microfluidic devices, biofabrication.