Application and advantages of high-throughput comet assay box COMPAC-50

High-throughput comet assay box COMPAC-50 --- A better comet experiment

Single-cell gel electrophoresis (comet assay) has been widely used as an experimental method for assessing DNA damage. From traditional institutions and research institutions to related pharmaceutical companies, more and more researchers have expressed strong interest in comet experiments. For example, screening of genotoxic drugs in pharmaceutical companies.

Figure 1 Basic comet experiment flow chart

Any kind of comet experiment must go through the above several experimental steps. However, low sample throughput and laborious operation are the main factors limiting the application of the experiment. In the traditional comet assay, the degree of DNA damage of 50 cells is measured one by one, which is quite time consuming. For the detection of DNA damage in large samples, Cleaver Scientific and the University of Leicester's oxidative stress team have jointly developed a new high-throughput comet electrophoresis system, COMPAC-50. The University of Leicester's Oxidative Stress Group detailed the upgrade process for this comet assay device in a paper published in the Scientific Report, mentioning the great effects of the new high-throughput glass shelves and high-throughput electrophoresis tanks on experiments. It also indicates that this upgrade greatly increases the number of samples to be analyzed, saves experiment time, reduces operating steps, and reduces the risk of reagent requirements and damage to slides. The compact design of the electrophoresis tank is very useful for laboratories with small bench space.

New method development materials used:

Simultaneous processing of 25 comet test slides using a POM carrier is known as a high-throughput glass frame (Figure 2A). This glass holder can hold the slides longitudinally and perform electrophoresis in the vertical direction. Earlier, the University of Leicester's oxidative stress team customized an electrophoresis tank (high-throughput electrophoresis tank 1; Figure 2B), which was suitable for demonstrating the experimental principle, but shortening the high-throughput glass frame to accommodate the electrophoresis tank. The electrophoresis cell has been redesigned to accommodate two high-throughput glass shelves (high-throughput electrophoresis tank 2, manufactured by Cleaver Scientific, UK) and can be used in subsequent experiments (Figure 2C, right).

Figure 2 (A) High-throughput glass shelf legend for 25 slides. The staining box can be used to perform lysis, neutralization, staining, and all cleaning-related steps. (B) High-throughput electrophoresis tank-1 and high-throughput glass shelves. The high-throughput electrophoresis tank-1 was originally used to demonstrate the experimental principle. (C) A difference diagram between the traditional comet experimental device (left) and the new high-throughput electrophoresis tank-2 (right), in which two devices share a single power supply.

This paper also investigates the effect of slide electrophoresis placed vertically on a high-flux glass shelf on the shape of the comet, the effect of buffer volume on the voltage and current of the electrophoresis cell, and the concentration stress and reproducibility of high-throughput comet assays. And other issues.

The results show that the results obtained with the upgraded version of the high-throughput comet electrophoresis system are not significantly different from the traditional ACA experiments, but the time saved is 60% , and the risk of gel damage is significantly reduced. In the matching of power supply devices, Cleaver Scientific's power supply (model: CS-300V) is also very suitable for this high-throughput comet experimental device in current and voltage configuration.

In later product development, Cleaver Scientific further improved the experimental setup, designed a suspended glass shelf (Figure 3A), and equipped a dye tray with a full set of high-throughput comet electrophoresis experiments for batch processing of glass. The film also has a ceramic cooling base (Figure 3B) and a cooling pack to ensure optimal electrophoresis buffer temperature and improved electrophoresis efficiency.

Figure 3 Suspension design of the glass frame and ceramic cooling base unit

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