User:Matt Hartings/Notebook/Photosynthesis/2012/02/10

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Extract crude K31C Ascaris hemoglobin from expression cells. Continued from here

Also, Tamra was in today and was working on extracting and transforming the DNA for sperm whale myoglobin that we received from Yi Lu. The vector that this contains this DNA is pT7-7. Tamra's notebook entry can be found here


I sonicated the cells (30 seconds in sonicator at power setting 11 followed by 30 seconds on ice, repeat procedure 2x)

After sonication, I evenly distributed the cells in centrifuge tubes (mass difference was on the order of 0.01g). These tubes were centrifuged for 2hours at 18000rpm and 4C. The red supernatant was poured into new storage tubes and then filtered to remove any extra particulates. After pouring off the supernatant, the color of the solution changed from pinkish to a darker red color indicating the oxidation of the heme) The filtered supernatant was placed in the refrigerator.


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