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A 2500 base-pair long cDNA of unknown sequence was cloned into the MCS of a plasmid vector that has universal priming sequences adjacent to and on both sides of the MCS. On one side this is called the "forward universal priming sequence" and on the other it is called the "reverse universal priming sequence." Sanger sequencing was done using primers that are specific for each of these universal priming sequences and approximately 700 nucleotides of sequence was determined from each primer. Assuming it is not possible to obtain sequence analyses longer than 700 nucleotides, describe the steps that must be done to obtain the remainder of the sequence of the cloned cDNA.

There are two very different methods that will accomplish this. One method requires much more effort and is less reliable than the other. Either method will receive full credit.

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  • Category:- Biology
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