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Hello students, to clone a fragment of human cftr gene into e.
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Coli using a plasmid vector with an ampicillin resistance gene and a multiple cloning site that is mcs within a lacz gene, follow these steps.
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First step is isolation of dna.
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Extract the human cftr gene from human cells using techniques like pcr or restriction enzyme digestion.
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Second one is plasmid preparation.
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Isolate a plasmid vector containing an ampicillin resistance gene and an mcs within a lacz gene.
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The lacz gene allows for blue or white screening.
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Third one is digestion with restriction enzymes.
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Cut both the isolated cftr gene fragment and the plasmid vector using the same restriction enzymes.
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This creates compatible ends for ligation.
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Moving to the ligation, mix the cut cftr gene fragment with the plasmid vector and use dna ligase to join them together.
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This creates a recombinant dna molecule.
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Then comes the transformation.
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Introduce the ligated dna into the e...