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bioRxiv · 10.1101/2023.06.26.546530

C2CAplus: a one-pot isothermal circle-to-circle DNA amplification system

Grasemann, L.·Pizarro, P. T.·Maerkl, S. J.
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Abstract

Rolling circle amplification (RCA) is a widely used DNA amplification method that uses circular template DNA as input and produces multimeric, linear single or double stranded DNA. Circle-to-circle amplification (C2CA) has further expanded this method by implementing product re-circularization using restriction and ligation, leading to a higher amplification yield, and enabling the generation of circular products. However, C2CA is a multistep, non-isothermal method, requiring multiple fluid manipulations and thereby compromises several advantages of RCA. Here, we improved C2CA to implement a one-pot, single step, isothermal reaction at temperatures ranging from 25 to 37{degrees}C. Our C2CAplus method is simple, robust, and produces large quantities of product DNA that can be seen with the naked eye.

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Grasemann, L., Pizarro, P. T., Maerkl, S. J.. 2023-06-26. C2CAplus: a one-pot isothermal circle-to-circle DNA amplification system. https://doi.org/10.1101/2023.06.26.546530

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