Tuesday, June 24, 2014

Cheyeanne Perez Day 6&7 Summary

               Yesterday I really didn't do anything that involved my project. I had helped create the LB media for growing more cells for a maxi prep (a lot more DNA is provided). We had to mix many powders together then autoclave it in a big machine on the second floor. That took a long time but after that ewe waited for it to cool down then added ampicillin to the LB media. Then we placed it in the shake incubator over night. Today we picked up where we left off and started a maxi prep. First we had to centrifuge all the cells in the LB media to get them to separate. We placed them in the centrifuge at least four times for fifteen minutes each. After each time we had to remove the unwanted media and dispose of it, which smelled really bad. Then  re-suspended the cells in the first solution and used a vortex machine to get rid of the clumps. Then we placed the second solution in the tubes and inverted the tubes eight times. Then we did the same for the third solution. Then we placed the tubes back in the centrifuge for thirty minutes. After that we removed the tubes and then  transferred the cells into another tube that had a filter in it and centrifuged it twice. Then we used sterile water to remove any unwanted proteins or cell parts. Then we placed the clean DNA in the freezer for any purpose we might use it for.

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