DNA is susceptible to be injured when attacked by free-radical (e.g. ·OH), which means desoxypentose was destroyed, phosphodiester bond was broken, basis group was broken and fallen off, then can further produce single strand breaks or double strand breaks. Take a small amount of cells immobilized in low melting point agarose to be coated on the slide glass, then destruct the cell membrane with alkali high salt solution and hydrolyze with alkali solution. Put the slide glass into the electrophoresis, NDA molecule will migrate to the positive pole under the effect of electric field. The electrophoresis speed will be fast if the DNA damaged severely and fragments were abundant. The un-damaged DNA macromolecule will be stranded in the original place due to the obstructing of the cell membrane. Staining with PI or EB, the cells with damaged DNA can be observed like comet, which can be used to making qualitative analysis, or quantitative analysis with corresponding software.
Kit Components (see Protocol for final kit quantities):Lysis Buffer, 100mLDMSO, 8mLNormal melting point agarose (NMA), 30mgLow melting point agarose (LMA), 30mgPropidium Iolide (PL), 400uL
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
