A terminal-block strategy for the amplified detection of DNA-break events

摘要: Abstract DNA break, including single-stranded break (SSB) and double-stranded (DSB), is the most serious damage. Sensitive detection of DNA-break events would be helpful for monitoring understanding as well screening preventing those chemical or physical damage factors. However, current methods suffer from some disadvantages complex, time-consuming, lowly sensitive especially only available SSB DSB only. In this work, we developed a novel terminal-block strategy convenient both events. Rolling circle amplification (RCA), well-known isothermal nucleic acid technique, was also introduced to improve sensitivity. our strategy, an oligonucleotide elaborately designed work primer RCA. As contrasted with conventional mono-functional primer, has three functional domains: reserved sequence at 5′-terminal hybridize circular probe, domain exposed inducements, blocking 3′-terminal block RCA reaction. The reaction can launched once cut off under break. Thus, signal translated into amplified readout. Fenton nicking endonuclease were adopted two model inducements in study. former induces random sites on DNA, whereas latter one specific site. Results show that these typical kinds analyzed using method, suggesting prospect universal application