Microwave Assisted Heating for the Rapid Sample Preparation of DNA for PCR Analysis
Author: Joanne Slatter
Publisher:
Published: 2013
Total Pages: 65
ISBN-13: 9781303462160
DOWNLOAD EBOOKBy using microwave radiation as a means to lyse bacterial cells, the extracted DNA can be amplified using the polymerase chain reaction (PCR). This work, in conjunction with the work of other collaborating labs on the project, allows for rapid detection of bacteria in a field portable device, termed microwave/prep-chip device. Cells will be introduced into the device via flow injection analysis and lysed by means of microwave radiation equipped with a flow cell. The lysate will then flow into a microfluidic device and be separated into its components based on charge. Results of these investigations have shown that DNA can be extracted from a whole cell sample suspended in a water solvent and amplified when the cells are exposed to microwave radiation at 90 °C for 5 minutes. After microwave heating lysing, DNA has been extracted and successfully amplified by adding the resulting supernatant to the PCR reaction mixture without additional sample clean up steps. It is important to determine conditions in which DNA can still be extracted and amplified, but also allow for the downstream separation and analysis of proteins, peptides, and lipids. The effects of an acidic solvent on DNA extraction and amplification were also tested between 0.1% and a 12% formic acid solvent, with successful amplification in the 0.1% to 4% formic acid solvent samples but no amplification in the 6% formic acid and greater samples. It has also been demonstrated that in increase in DNA amplification can be seen when the whole cell samples are microwave heated in a gold nanoparticle solution.
