The following is a list of the various steps involved in the cloning of a DNA fragment from human cells into a plasmid v

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The Following Is A List Of The Various Steps Involved In The Cloning Of A Dna Fragment From Human Cells Into A Plasmid V 1 (56.62 KiB) Viewed 14 times

The following is a list of the various steps involved in the cloning of a DNA fragment from human cells into a plasmid vector. The vector contains genes for both tetracycline resistance and ampicillin resistance. The vector contains only one site where restriction enzyme "X" can cut and this site is located in the ampicillin resistance gene. The order in which the different steps (a toj) are provided is incorrect. Indicate the correct order. (a) Isolate plasmid DNA from bacteria obtained from a single colony that can grow on tetracycline but not on ampicillin plates. Join cut plasmid DNA to human DNA fragments with the ligase enzyme. (b) (c) Select bacterial colonies that are unable to grow on ampicillin. (d) Transform bacterial cells with recombinant plasmid. (e) Determine the size of the cloned DNA fragment. (1) Isolate DNA from a human cell line. (8) Isolate plasmid vector DNA from bacterial cells and cut with restriction enzyme (h) Select bacterial cells that are able to grow on tetracycline plates. () Compare the recombinant plasmid with the vector plasmid by means of electrophoresis after cutting both with restriction enzyme "X". 0) Cut human DNA with restriction enzyme "X"