The correct answer is Option (2) → (C), (D), (A), (B)
In recombinant DNA (rDNA) technology, the steps occur in a specific sequence to ensure that the desired genetic material is accurately isolated, modified, and introduced into a host cell. Here’s an explanation of each step in the correct order:
- Isolation of the genetic material (Step C): The process begins by extracting the DNA containing the gene of interest from a cell. This is often done by breaking open the cells and using techniques to separate the DNA from other cellular materials.
- Cutting the DNA at specific locations (Step D): Once isolated, the DNA is cut at specific points using restriction enzymes. After having cut the source DNA as well as the vector DNA with a specific restriction enzyme, the cut out ‘gene of interest’ from the source DNA and the cut vector with space are mixed and ligase is added. This results in the preparation of recombinant DNA.
- Amplification of gene by PCR (Step A): After cutting, the gene of interest is amplified using PCR (Polymerase Chain Reaction). This technique makes multiple copies of the gene, ensuring there is enough DNA available for the next steps in the rDNA process.
- Insertion of rDNA into host cell using vector (Step B): After the DNA fragment is ligated into the vector, the recombinant DNA is ready for transfer into the host organism, such as bacteria, plant cells, or animal cells. This transfer is usually achieved through transformation methods specific to the host organism, such as electroporation or biolistics.
This order (C, D, A, B) ensures that the gene is accurately isolated, prepared, and effectively introduced into a host, where it can be studied or used for further applications.
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