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Bio383H Lab 1 Results p. 2 draft

Submitted by liamharvey on Thu, 02/01/2018 - 19:23

The purpose of this experiment was to extract, purify and quantify the DNA of Brachypodium distachyon a common grass species. The extraction and purification process involved three steps; breaking up tissue and cells, preventing DNA degradation and removing unwanted molecules. To break up the tissue and cells of B. distachyon, mechanical grinding was first performed using a ball mill to break up the extracellular matrix. Chemical disruption was performed next using the detergent sodium dodecyl sulfate (SDS) with metal chelation using ethylene diamine tetra-acetic acid (EDTA). The final step in breaking up tissues and cells was heat disruption to release the DNA which was done on a hot bock set to 65°C. After DNA had been extracted from B. distachyon tissue, it was essential to prevent the enzyme DNase from degrading the free DNA. Preventing DNase from breaking down the extracted DNA was done with the already added EDTA. DNase requires Mg++ as a cofactor; by using EDTA to chelate with Mg++ prevents DNase from acting.