Does it matter if the same pH is used for alkali unwinding and alkaline electrophoresis?
The same solution (> pH 13) is used for alkali unwinding and alkaline electrophoresis. To avoid variation due to pH, the buffers should be prepared fresh. At pH 12.1, initial breaks are analyzed, while at pH 12.5 and pH 13 alkaline labile adducts are converted to breaks. At pH 12.5, abasic lesions are converted to single strand breaks and at pH 13, additional labile sites are converted to single and double strand breaks. Maximum damage caused by an agent is visualized at pH 13 in CometAssay and FLARE Assay.
