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dna extraction

Satisfactory Essays
November 5, 2014
Bioinformatics
Leigh Ann Santana
DNA Extraction Lab DNA extraction is an important process because the DNA first needs to be purified away from proteins and other cellular contaminants. Cell are needed, because that is where the DNA is located. Inside almost every cell in our bodies is a nucleus, and inside each nucleus is about two meters of DNA. The following steps are needed to purify DNA from a cheek swab. Collect cheek cells, Burst cells open to release DNA, separate DNA from proteins and debris, and isolate concentrated DNA. The end of the swab must be cut off so it will be able to close in the tube. Using the micropipette, add some lysis solution to the tube. Lysis is a Greek work that means “to separate”. The lysis solution added contains two important ingredients: detergent and an enzyme call proteinase K. The detergent disrupts the cell membrane and nuclear envelope, causing the cells to burst open and release their DNA. The DNA is still wrapped very tightly around proteins called histone, and the proteinase K cuts apart the histones to free the DNA. Now the warm water bath is needed. The cells have to stay in the warm water bath long enough for the DNA to be freed from the cells, and the swab is removed from the tube. Add concentrated salt solution to the tube. The salt caused proteins and other cellular debris to clump together. In order to “balance” the centrifuge, with a tube containing water is placed opposite the DNA tube. Close the lid and turn it on. Inside the centrifuge, the tubes spin around at high speed. The heavy clumps of protein and cellular debris to sink to the bottom of the tube, while the strands of DNA remain distributed through the liquid. Now use the micropipette to carefully remove the top liquid (which contains DNA) and place the liquid into a clean tube. The proteins and other cellular debris stay behind. Now add isopropyl alcohol to the tube. Inverting the tube several times mixed the isopropyl alcohol into the DNA solution. Because DNA is not soluble in isopropyl alcohol, it comes out of the solution. The clumped DNA can be seen without a microscope. Place the tube into the centrifuge and this time after the sample spins the DNA sinks to the bottom of the tube. One the liquid is removed and the DNA is allowed to dry, the solution of your choice the DNA can re-dissolve. The DNA can be frozen for up to several years.

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