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Dystrophin is a huge gene (25,000,000 bases). As such, chain termination sequenc

ID: 55107 • Letter: D

Question

Dystrophin is a huge gene (25,000,000 bases). As such, chain termination sequencing must be done in many parts. Assuming that you don’t know anything about the sequence to begin with, how can you amplify the sequence for sequencing?


      
Put the cut sequences into a plasmid for which you know some sequence. Then, complete PCR.

      
Put the cut sequences into a plasmid. Then transform the plasmids into bacteria and allow them to replicate.

      
Both (a) and (b) can be used.

      
Neither (a) nor (b) can be used.

Explanation / Answer

Neither (a) nor (b) can be used.

The statement given points towards a method of sequencing known as primer walking which itself is based on primers for PCR. To carry out PCR something should be known about the sequence prior to the method so that primers can be designed accordingly.