MALCOLM BUCKLE

Over a long period I have developed an approach using the probability of UV induced cyclobutane dimer formation between adjacent pyrimidines on the same DNA chain to follow changes in local base structure in DNA and nucleoprotein complexes. This technique called Photo-chemical Analysis of Structural Transitions (PhAST) promotes DNA pyrimidine dimer formation. The location and intensities of this photo-chemistry are affected by external agents that reshape the DNA structure. Thus, the technique provides unique information on local structure at the DNA nucleotide level. We reveal photochemical products using a simple primer extension technique coupled to capillary electrophoresis (now being extended to NextGen sequencing!). Our approach confers high-resolution, excellent quantification, application in vitro and in vivo, and the possibility of high-throughput since practically any size DNA sample may be analysed.