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

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

Biology, Academics

  • Category:- Biology
  • Reference No.:- M937116

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