Phosphorothioate-based ligase-independent cloning

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Phosphotothioate-based ligase-independent cloning^[1] is a cloning method that uses the Phosphotothioate chemistry as a new universal cloning starting with the target's gene and vector's amplification with PCR. It is also commonly known as the PLICing method and it consists of three main steps: amplification, cleavage and hybridization. This method is important because it is a sequence-independent method to clone DNA fragments in an easier way than the traditional way which requires the use of restriction enzymes for DNA ligase, and it is also a really high efficient method.^[2]