Cold Spring Harb Protoc. 2020 Apr 1;2020(4):098061. doi: 10.1101/pdb.prot098061.
Plasmid DNA is prepared from the recombinant shuttle vector pLD53.SCAB/A-B created by cloning of the A and B homology arms for two-step bacterial artificial chromosome (BAC) engineering. To confirm that the A-box and B-box arms have been successfully incorporated into pLD53.SCAB, the pattern of enzyme digestion of the modified plasmid is compared with that of the unmodified pLD53.SCAB. Once the shuttle vector is shown to carry the proper sequences, it is ready for transfer into the BAC host.
质粒 DNA 是从重组穿梭载体 pLD53.SCAB/A-B 中制备的，该载体是通过克隆两步法细菌人工染色体 (BAC) 工程的 A 和 B 同源臂而构建的。为了确认 A 框和 B 框臂已成功整合到 pLD53.SCAB 中，需要比较修饰后的质粒的酶切图谱与未修饰的 pLD53.SCAB 的酶切图谱。一旦证明穿梭载体携带了适当的序列，就可以准备将其转移到 BAC 宿主中。
