| 摘要 |
Described is a method for preparing a nucleic acid for sequencing. The method comprises a) amplifying a nucleic acid template using a dNTP mix that contain 5-methyl dCTP, thereby producing product nucleic acid molecules that contain methylcytosines; b) digesting the product nucleic acid molecules with a MspJI family restriction endonuclease, thereby cleaving the product nucleic acid molecules at sites that are adjacent to at least some of the methylcytosine, and producing fragments of the product nucleic acid molecules; and c) ligating double-stranded adaptors onto the ends of the fragments to produce adaptor-ligated products. The enzymes used include MspJI, FspEI, LpnPI and SgeI. The amplification method may be PCR or isothermal amplification. Also described is a kit for carrying out the method including a dNTP mix containing 5-methyl dCTP, one or more MspJI-family restriction endonucleases, and double-stranded adaptors. |
