To remove DNA from a eukaryotic sample, the first step is cracking open the cell's various membranes to get to the nucleus, where the DNA is stored. Lysis buffers are solutions of chemicals that break down the cellular membranes of a sample and also cause proteins to precipitate out of solution. Lysis buffers are generally alkaline and contain detergents, chemicals with both hydrophobic and hydrophilic portions of the molecule. These detergents and the alkalinity of the solutions cause the cellular membranes to degrade by disrupting the hydrophobic membranes. Other chemicals can be added to the lysis buffer to regulate pH and to cause unwanted macro-molecules, such as proteins, to precipitate.
So this week in lab I made my lysis buffer. None of the commercially available buffers had the same concentrations of chemicals as the protocol I'm following. I had to make 10mM Tris, 10mM EDTA, 2% SDS pH 8, from more concentrated chemicals found in the lab. It was great to practice dilution again. Diluting chemicals is a precise method that I feel is not one of my strengths and thus something I must practice more.
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