Difference between revisions of "20.109(F16):Data analysis (Day7)"
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===Part 1: Visualize H2AX assay results=== | ===Part 1: Visualize H2AX assay results=== | ||
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#Aspirate the secondary antibody solution. | #Aspirate the secondary antibody solution. | ||
#To wash the coverslips, add ~500 μL of TBS per well and rock the plate back and forth, then aspirate. | #To wash the coverslips, add ~500 μL of TBS per well and rock the plate back and forth, then aspirate. | ||
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#Complete Steps #5-7 for coverslips from two gamma-irradiated and two control wells. | #Complete Steps #5-7 for coverslips from two gamma-irradiated and two control wells. | ||
#Alert the teaching faculty when all four microscope slides are ready and you will be escorted to the microscope in the Engelward laboratory. | #Alert the teaching faculty when all four microscope slides are ready and you will be escorted to the microscope in the Engelward laboratory. | ||
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| + | ===Part 2: Data analysis for H2AX assay=== | ||
==Reagents== | ==Reagents== | ||
Revision as of 18:57, 25 August 2016
Contents
Introduction
Protocols
Part 1: Visualize H2AX assay results
- Aspirate the secondary antibody solution.
- To wash the coverslips, add ~500 μL of TBS per well and rock the plate back and forth, then aspirate.
- Repeat this step a total of two times.
- Add 500 μL TBS per well.
- Obtain a fine gauge (26 3/8) needle, a pair of tweezers, and four glass microscope slides from the front laboratory bench.
- Carefully press the tip of the needle against the benchtop to bend it into a right angle such that the beveled side of the needle is the interior angle.
- Use the 'hook' created in Step #4 to lift the coverslip from the bottom of the well, then use the tweezers to 'catch' the coverslip.
- In total, you only need to visualize two coverslips from the gamma-irradiated plate and two coverslips from the control plate. Therefore, you have several wells that can be used as practice.
- When you are confident with your ability to retrieve the coverslips from the wells, add 5 μL of mounting media to a glass microscope slide.
- Place the coverslip cell-side down on the mounting media on the microscope slide.
- The cell-side of the coverslide is the side that was facing up in the well of the plate.
- Complete Steps #5-7 for coverslips from two gamma-irradiated and two control wells.
- Alert the teaching faculty when all four microscope slides are ready and you will be escorted to the microscope in the Engelward laboratory.
Part 2: Data analysis for H2AX assay
Reagents
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