DNA Isolation: Experimental reagents

You are a final-year undergraduate student completing your thesis project in the laboratory of Dr. Demetriou. Dr. Demetrious’s work focuses on elucidating the pathophysiological mechanisms contributing to the development of heart failure. The work hopes that these insights will provide novel therapeutic targets. Your project focuses on studying the role of the newly-identified protein noveltin in cardiac function and dysfunction, using a transgenic mouse model. Your work will entail comparative experiments with control mice, which over-express the noveltin gene. 

It is very important that mice carrying the gene (experimental group) are appropriately identified and compared to mice with normal expression of noveltin (control group).  Your supervisor asks you to first isolate DNA from mouse tails and then carry out a polymerase chain reaction (PCR) to genotype the mice. 

You decide to use a DNA purification kit (QIAGEN), which is commonly used in your supervisor’s laboratory. 

Website: DNA purification kit (QIAGEN)

You look at the manufacturer’s instructions and note that a number of reagents are used including lysis buffers, proteinase K, ethanol, wash buffers, and an elution buffer. While the steps are clearly outlined, there is no information to explain the role of each reagent in the kit so you discuss this with your supervisor and he asks you which reagent is primarily responsible for denaturing proteins and aiding in the breakdown of tissues and cells.