Cold Spring Harb Protoc. 2021 Apr 1;2021(4):2021/4/pdb.prot101279. doi: 10.1101/pdb.prot101279.
To generate polymerase chain reaction (PCR) products that can be directionally cloned into a vector, different restriction sites are built into the forward and reverse primers that are used in the PCR. After PCR, the amplified product is purified, cleaved with the appropriate restriction enzymes, ligated into a vector with compatible cohesive ends, and used to transform .
为了生成可定向克隆到载体中的聚合酶链反应 (PCR) 产物，正向和反向引物中构建了不同的限制酶切位点，这些引物用于 PCR。PCR 后，扩增产物被纯化，用适当的限制酶切割，连接到具有互补粘性末端的载体中，并用于转化。
