Evaluation of primary DNA-damage is one way to identify potential genotoxic agents and for this purpose the Comet assay has, for the last decades, been used to monitor DNA single strand and double strand breaks in individual cells. Various attempts have been made to modify the different steps in the in vitro protocol for the Comet assay in order to improve its sensitivity. However, to the best of our knowledge, nobody has tried to replace the traditionally used NaOH-based electrophoresis solution (pH > 13), with another type of solution. In the present paper, using TK-6 cells exposed to different concentrations of H₂O₂ or ionizing radiation, we present evidence clearly showing that a low-conductive LiOH-based electrophoresis solution at pH 12.5, and a more gentle lysis procedure, significantly improved both the speed and sensitivity of the assay. The new approach, which we call the Flash-comet, is based on a lysis buffer at pH 8.5, an unwinding time of 2.5 min in a LiOH solution without EDTA at pH 12.5, and an electrophoresis time of 1 min at 150 V (5 V/cm) using the same solution.

评估初级 DNA 损伤是识别潜在基因毒性物质的一种方法，为此目的，彗星试验在过去几十年中一直用于监测单个细胞中的 DNA 单链和双链断裂。已经进行了各种尝试来修改彗星试验体外方案中的不同步骤，以提高其灵敏度。然而，据我们所知，没有人尝试用另一种类型的溶液代替传统使用的基于 NaOH 的电泳溶液（pH > 13）。在本文中，使用暴露于不同浓度 H ₂ O _{2 的}TK-6 细胞或电离辐射，我们提供的证据清楚地表明，pH 12.5 的低导电性 LiOH 基电泳溶液和更温和的裂解过程，显着提高了测定的速度和灵敏度。我们称之为 Flash-comet 的新方法基于 pH 8.5 的裂解缓冲液，在 pH 12.5 的不含 EDTA 的 LiOH 溶液中解旋时间为 2.5 分钟，以及在 150 V (5 V/cm) 使用相同的溶液。