Question

Illustrate steps to show the construction of a recombinant DNA molecule.

Hint:

Remember the key steps involved in making a recombinant DNA molecule. Understand the role of restriction enzymes, DNA ligase and host organisms in this.

Answer 1

Recombinant DNA molecule construction typically entails these stages: DNA Isolation: Obtain the desired DNA fragment and a vector DNA (often a plasmid) destined for host organism introduction. DNA Cleavage: Employ the identical restriction enzyme to cut both the target DNA and vector DNA, generating compatible ends for subsequent joining. Ligation: Utilize DNA ligase to covalently link the target and vector DNAs, forming a recombinant DNA molecule. Transformation: Introduce this recombinant molecule into a host organism, such as E. coli, enabling its replication along with the inserted DNA. Selection and Screening: Assess transformed cells to confirm successful insertion, commonly through antibiotic resistance assays.