Boyer and Cohen invented the recombinant DNA method in 1973.
It's a technique for creating an artificial DNA molecule by merging two or more DNA fragments that aren't necessarily related to one another.
Steps to produce recombinant DNA:
DNA Isolation:
DNA is isolated in its purest form, meaning there are no other macromolecules present.
DNA must be separated and purified using enzymes such as lysozymes, cellulase, chitinase, ribonuclease, and proteases since it coexists with other macromolecules such as RNA, polysaccharides, proteins, and lipids within the cell membrane.
DNA splicing:
Restriction enzymes are essential for this phase. It aids in determining the place in a vector genome where a specific gene is inserted. Restriction enzyme digestions are the name for this reaction.
DNA amplification:
PCR or polymerase chain reaction is used to amplify copies of genes. After restriction enzymes have cut the desired gene of interest, it is simply a process of multiplying a single DNA copy into multiple copies.
It permits a single copy or a few copies of DNA to be multiplied by thousands or millions.
Bringing DNA together:
In this stage, the vector and a piece of DNA are linked together. The enzyme DNA ligase is used to do this.
The resulting DNA molecule is a cross between the interest and vector DNA molecules. The merging of distinct DNA strands is referred to as recombination in genetics.
As a result, the new hybrid DNA molecule is referred to as a recombinant DNA molecule, and the technique is referred to as recombinant DNA technology.
rDNA insertion into a host:
In this stage, the recombinant DNA is transported into a recipient host cell, most typically a bacterial cell. 'Transformation' is the word for this operation.
