A. After you performed the freeze-thaw cycle to lyse the bacteria, you pelleted the debris using...
A. After you performed the freeze-thaw cycle to lyse the bacteria, you pelleted the debris using a centrifuge.What two reasons did this pellet fluoresce green?(Yes, I know it contained GFP…. but why?)
B. <w1 > should not have fluoresced. Why? (I know it did not contain rGFP, but why?) W2-W4 should have fluoresced slightly. Give two possible reasons why. (Yes, I know it contained GFP…. but why?)
Homework Answers
1. During freeze-thaw cycle the cell will lyse and debris will flot on the solution it contain GFP. The GFP molecules binds to the debris now after centrifuge the pellets contain debris with GFP so it emits the fluroscence.
2. W1 contain only breaking buffer so it will remove all the GFP from the column. W2-W4 have fluroscence because His tag not bound to Ni2+ agarose beads and it will washed away or the beads will saturated with GFP and excess was washed away.
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A. After you performed the freeze-thaw cycle to lyse the
bacteria, you pelleted the debris using...
After you performed the freeze-thaw cycle to lyse the bacteria, you pelleted the debris using a...
After you performed the freeze-thaw cycle to lyse the bacteria, you pelleted the debris using a centrifuge. What two reasons did this pellet fluoresce green? (Yes, I know it contained GFP…. but why?)
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