| 摘要 |
PURPOSE:To provide a method of preparing a ligated double-stranded DNA which can freely, easily and specifically ligate a DNA fragment, by freely selecting from the base sequence and the base number of the cohesive end with reduced or no restriction on the sites to be cleaved by a restriction enzyme. CONSTITUTION:Using a primer, as at least one side of the primer DNA, which contains ribonucleotide groups in which the sequence on the 3' side of the ribonucleotide group is at least partially complementary to the end part of the area to be magnified of the plus or minus chain of the template DNA and the sequence on the 5'-side of the ribonucleotide group is that for forming the cohesive end, the polymerase chain reactions (PCR) is performed to magnify and synthesize a double-stranded DNA fragment. Further, fragment is treated with ribonuclease or an alkali, phosphorylated to give a double-stranded DNA fragment having a cohesive end. Then, this double-stranded DNA fragment is ligated with another double-stranded DNA fragment which is complementary to this fragment using a DNA ligase. |
