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DNA Spooling - Westminster College

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<strong>DNA</strong> <strong>Spooling</strong>improve the accuracy of this measurement.) Then determine the appropriate volume ofchilled ethanol required for <strong>DNA</strong> precipitation.9. Slowly layer the appropriate amount of chilled ethanol on top of the homogenate. Alayering effect can be achieved by tilting the beaker containing the cell homogenate andthen slowly pouring the ethanol down the side of the beaker. White strands ofprecipitated <strong>DNA</strong> may appear.Figure 1. Layering of Ethanol on Homogenatecold ethanolhomogenate10. Using a glass stirring rod,wind the <strong>DNA</strong> around the rod byslowly spinning the glass rodwhile stirring the mixture in onedirection. Strands of precipitated<strong>DNA</strong> should begin to appear atthe interface of the cellhomogenate/ethanol mixture.Continue stirring and collectingprecipitated <strong>DNA</strong> for a fewminutes.11. Once the <strong>DNA</strong> is collected,gently scrape it into a 1.5 mLmicro centrifuge tube that contains 1 mL of water or low salt buffer (e.g. TE). If storedin a refrigerator, the <strong>DNA</strong> sample can last for several months and be used for otherexperiments.<strong>Westminster</strong> <strong>College</strong> SIM 3

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