Difference between revisions of "20.109(S14):DNA repair assays(Day6)"
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===Part 1: Prepare cells for flow cytometry=== | ===Part 1: Prepare cells for flow cytometry=== | ||
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+ | #Begin by briefly looking at your cells under the microscope. Do the cells in any wells appear less dense or less healthy than in others? Note down any such observations. | ||
+ | #Aspirate the media from each well according to the protocol below. | ||
+ | #*As you work, tip the plate down a little to pool the media at the bottom of each well. | ||
+ | #*Place your aspirator at the bottom of a well, and suck up the media. Remove all the liquid, but remove the aspirator promptly after that or you may damage/aspirate some cells. | ||
+ | #*Before moving to the next (non-duplicate) well, dip your aspirator briefly (less than a second!) in ethanol. Then hold it up to dry and count to 3. | ||
+ | #*Distribute 0.5 mL of warm PBS to each well, using the 2 mL pipet. | ||
+ | #* | ||
===Part 2: Plate irradiated cells for inhibitor dose response=== | ===Part 2: Plate irradiated cells for inhibitor dose response=== |
Revision as of 16:45, 25 March 2014
Contents
Introduction
Protocols
Part 1: Prepare cells for flow cytometry
- Begin by briefly looking at your cells under the microscope. Do the cells in any wells appear less dense or less healthy than in others? Note down any such observations.
- Aspirate the media from each well according to the protocol below.
- As you work, tip the plate down a little to pool the media at the bottom of each well.
- Place your aspirator at the bottom of a well, and suck up the media. Remove all the liquid, but remove the aspirator promptly after that or you may damage/aspirate some cells.
- Before moving to the next (non-duplicate) well, dip your aspirator briefly (less than a second!) in ethanol. Then hold it up to dry and count to 3.
- Distribute 0.5 mL of warm PBS to each well, using the 2 mL pipet.
Part 2: Plate irradiated cells for inhibitor dose response
For next time
Reagent list
write something here or not accessible to edit
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