20.109(S14):DNA repair assays(Day6)

Introduction

Protocols

Part 1: Prepare cells for flow cytometry

1. 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.
2. 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

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