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7 September, 23:45

1. List each of the seven steps involved in rDNA technology, from obtaining large amounts of this gene to confirming your pure protein X gene from an E. coli and describe one of them briefly (2 pts).

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  1. 7 September, 23:54
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    Recombinant DNA technology can be described as a technology in which DNA of two different organisms is merged and put into a host. This technology allows the production of genes which are beneficial to humans.

    The steps of the rDNA technology are:

    1) Isolating of DNA fro a cell

    2) Using Restriction Enzymes to cut the DNA

    3) Isolation of the desired DNA

    4) Amplification of the desired gene

    5) Ligating the DNA fragment into a suitable vector

    6) Transfer of the foreign DNA into an host organism

    7) Getting the gene product

    One of the process to describe will be:

    Amplification of the desired gene: The most common method of amplifying a gene in vitro is by the technique of PCR.

    There are three basic steps of PCR:

    Denaturation:

    In this step, the two strands of the DNA are separated by heat. Usually the temperature is set to 92-95 degrees for 15 sec Celsius for this process.

    Annealing:

    In this step, primers are used to anneal the DNA strands. This step is carried out at lower temperatures mainly in between 60-65 degrees Celsius.

    Extension: The Taq polymerase enzyme adds nucleotides to the primers complementary to the DNA strands.
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