Genomic_DNA_from_flies

Genomic_DNA_from_flies - Isolation of genomic DNA from...

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Isolation of genomic DNA from flies February 9, 2009 There are kits available for the isolation of DNA from tissue but for large amounts, I prefer this old- fashioned technique. It is a four stage technique, during which 1) a cell suspension is prepared from tissue, 2) the cells are lysed with SDS and proteins are degraded with proteinase K, 3) the lysate is extracted with phenol and chloroform, and 4) the DNA is precipitated with salt and ethanol. General procedure: 1. Collect about 200 or so fruit flies in an empty container. Let them sit for a few hours and then freeze overnight . The hours of sitting allows anything they have eaten to be digested. Therefore, there will be no contaminating DNA from yeast, for example. 2. Pre-chill a mortar and pestle on ice. Add the flies and grind away until you generate a thick paste. 3. Add 4 ml of homogenization buffer and resuspend the paste. Carefully pour the paste into a pre-chilled Dounce homogenization tube. Rinse the mortar with a few mls of buffer and also add that to the tube.
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This note was uploaded on 11/25/2009 for the course BI 379 taught by Professor Kavaler during the Spring '09 term at Colby.

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Genomic_DNA_from_flies - Isolation of genomic DNA from...

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