Difference between revisions of "Team:UCLA/Notebook/Recombinant Expression/7 July 2015"
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'''Lysis Protocol''' | '''Lysis Protocol''' | ||
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1. Add Lysozyme. | 1. Add Lysozyme. | ||
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2. Incubate on ice for 30 minutes. | 2. Incubate on ice for 30 minutes. | ||
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3. Add PMSF solution, and while swirling, slowly add Deoxycholic Acid in powder form. | 3. Add PMSF solution, and while swirling, slowly add Deoxycholic Acid in powder form. | ||
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4. Incubate at 37 degrees Celsius for 20 minutes. | 4. Incubate at 37 degrees Celsius for 20 minutes. | ||
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5. Add DNAse. | 5. Add DNAse. | ||
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6. Incubate on rocking incubator at room temperature for 30 minutes. | 6. Incubate on rocking incubator at room temperature for 30 minutes. | ||
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7. Centrifuge sample for 15 minutes at 3,000g at 4 degrees Celsius. | 7. Centrifuge sample for 15 minutes at 3,000g at 4 degrees Celsius. | ||
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8. Collect supernatant and store at -20 degrees Celsius. | 8. Collect supernatant and store at -20 degrees Celsius. | ||
Revision as of 00:39, 10 July 2015
Lysis of Cells, Testing the Protocol with Fractions 1-5
Lysis Protocol
1. Add Lysozyme.
2. Incubate on ice for 30 minutes.
3. Add PMSF solution, and while swirling, slowly add Deoxycholic Acid in powder form.
4. Incubate at 37 degrees Celsius for 20 minutes.
5. Add DNAse.
6. Incubate on rocking incubator at room temperature for 30 minutes.
7. Centrifuge sample for 15 minutes at 3,000g at 4 degrees Celsius.
8. Collect supernatant and store at -20 degrees Celsius.