| 主权项 |
1. A method of amplifying at least two different nucleic acid sequences, the method comprising:
a) creating known 5′ and 3′ ends of at least two nucleic acid sequences by restriction enzyme digestion, b) combining, in a single reaction, a plurality of upstream and downstream nucleic acid patches, a plurality of upstream and downstream universal primers, and the nucleic acid sequences of known ends from step (a), wherein the downstream universal primers comprise a protecting group; c) performing at least two annealing cycles, wherein each cycle comprises denaturing the nucleic acid sequences of known ends from step (a), and cooling the reaction to anneal an upstream and downstream nucleic acid patch to a nucleic acid sequence of step (a) and an upstream universal primer to an upstream patch and a downstream universal primer to a downstream patch; d) ligating the upstream universal primer and the downstream universal primer to each nucleic acid sequence; and e) amplifying the nucleic acid sequences of step (d). |
