Research

Research Areas: Highlights    |    Detecting Sequence Change in vivo    |    High Throughput DNA Damage Quantification    |    Research Overview

New Technologies: Microarray for High Throughput DNA Damage Quantification

two comets

Increasing amounts of DNA damage are visible in the comets below. The nucleoid from a normal cell is on the left, whereas the nucleoid from a cell exposed to DNA damage is on the right.

stampFor decades, researchers have measured DNA damage using the single cell gel electrophoresis assay, or comet assay. Originally developed by Singh et al in 1988, the basis for the assay is that damaged DNA migrates more readily than undamaged DNA when electrophoresed. Although effectively used in thousands of publications, the assay nevertheless suffers from sample-to-sample variation and it is highly laborious.

To increase reproducibility and throughput, we collaborated with the Bhatia laboratory (MIT HST EECS). Using microfabrication, we developed a method for making an array of microwells in agarose. A single cell (or groups of cells) can be loaded into each well.

science image  comet grid

  • Increased Throughput
  • Permits a broader range of cell concentrations
  • High consistency and efficiency
  • Lower inter- and intra- experimental noise
  • Detects many classes of DNA lesions
  • Detects multiple repair pathways
  • Can be used to measure repair capacity in human cells

 

Application of the Comet Microarray

Analysis of inter-individual variation in DNA repair capacity.

     

Studies of exposure-induced DNA damage and modulation of repair.

Research image       image composite
         
 

High throughput screening for optimization of chemotherapy.

   
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