Identification of Pet14b Bgal Plasmid Transformation in E coli Dh5a Cells Using PCR Technique
Sukmawati Sukmawati, Iksan Badaruddin, Fatimah Hardianti, Erin D Noya, Cristy Radjawane, Ayusal Salam

Universitas Muhammadiyah Sorong

Abstract

Abstract. DNA transformation is one method for inserting recombinant DNA into bacterial cells. This method is now widely used to transfer plasmids from one bacterial strain to another.The purpose of this study was to identify the results of PET14b-&#946-gal plasmid transformation into E. coli DH5&#945- cells using PCR technique. The method used in this study is the colony pcr technique. The results of this study indicate that the locus is well amplified, namely the emergence of the tape which all smears. In the forward (sense) area of the band length of 600 bp, an increase in size indicates that the orientation is correct, whereas in the reverse area (antisense) the band length of 400 bp indicates the orientation is reversed. This shows that the DNA sample used contains a number of DNA sequences that complement the primer. It was concluded that Plasmid pET14b is 600 bp in length in the forward direction and the &#946- galactosidase gene inserts antisense at the cloning site in the lacZ gene region.

Keywords: PET14b-&#946-GAL plasmid, Transformation, E. Coli DH5&#945-, PCR technique

Topic: Physics, Biology, and Chemistry