Abstract
Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method that amplifies DNA with high specificity, efficiency, and rapidity under isothermal conditions (1). The LAMP method requires a set of four specially designed primers and a DNA polymerase with strand displacement activity. The amplification products are stem-loop DNA structures with several inverted repeats of the target and cauliflower-like structures with multiple loops, yielding >500 μg/mL. Although LAMP amplifies DNA under isothermal conditions, the template DNA is heat-denatured. To determine whether LAMP can be performed under isothermal conditions at all steps, we attempted to amplify DNA without using a heat-denatured template. Hepatitis B virus (HBV) DNA that was obtained from a patient was digested with Bam HI and cloned into pBR322 plasmid vector. Plasmid and genomic DNA was prepared using the plasmid Midi reagent set (Qiagen) and EXTRAGEN reagent …
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