Simplified plasmid cloning with a universal MCS design and bacterial in vivo assembly.

ABSTRACT:

Background

The ability to clone DNA sequences quickly and precisely into plasmids is essential for molecular biology studies. The recent development of seamless cloning technologies has made significant improvements in plasmid construction, but simple and reliable tools are always desirable for time- and labor-saving purposes.

Results

We developed and standardized a plasmid cloning protocol based on a universal MCS (Multiple Cloning Site) design and bacterial in vivo assembly. With this method, the vector is linearized first by PCR (Polymerase Chain Reaction) or restriction digestion. Then a small amount (10 ~ 20 ng) of this linear vector can be mixed with a PCR-amplified insert (5× molar ratio against vector) and transformed directly into competent E. coli cells to obtain t

SUBMITTER: Chen F 

PROVIDER: S-EPMC7962268 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature