20.109(F16):Asses DNA repair variability with comet chip (Day4)

Introduction

Protocols

Part 1: Load cells and induce DNA damage

For this experiment everyone will use H₂O₂ to induce DNA damage in three Coriell cell lines to assess the DNA repair capacity variability in healthy individuals. Use the following plate map to as you complete the below steps:

Plate map for comet chip repair assay. The labels on the right refer to the Coriell cell lines and the labels on the bottom refer to the H₂O₂ dose. Grey wells will be empty for technical reasons.

You will load comet chips prepared by the teaching faculty using the procedure from M1D2; however, please note the following differences:

• You will not complete the lysis incubation immediately following the cell loading, rather this will be done after the DNA damage induction step.
• For the cell number you will use 500 K/mL and for the loading time you will use 30 min.

Part 2: Use comet chip to assess repair capacity

Reagents

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