Abstract
Nicking enzymes are a special family of restriction endonucleases, which recognize a specific sequence along double-stranded DNA, but cut only one DNA strand, thus creating a DNA nick in the corresponding strand. The nick can be employed in the rolling circle amplification (RCA) reaction, which has attracted substantial interest lately due to its high sensitivity, excellent specificity, and detection versatility. We have recently developed a new nonlinear RCA format by introducing a nicking enzyme into the hyperbranched RCA system. This innovative DNA amplification technique, named as netlike RCA (NRCA) and described in this chapter, resulted in cubic amplification of a HIV-1 DNA target sequence and strongly discriminated it from its single-base mutants.
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