Biology Practice Problems PCR and Dideoxy Sequencing Practice Problems Solution: You are interested in the sequence of a 10-base se...

Solution: You are interested in the sequence of a 10-base segment f a gene. Using the chain-terminator procedure, you obtain the results shown below after gel electrophoresis.A. What is the sequence read from the gel (5'→3')?B. What is the sequence of the gene segments (5'→3')?

Problem

You are interested in the sequence of a 10-base segment f a gene. Using the chain-terminator procedure, you obtain the results shown below after gel electrophoresis.

A. What is the sequence read from the gel (5'→3')?

B. What is the sequence of the gene segments (5'→3')?

Solution

The Sanger method, or the chain-terminator method determines the sequence of the template strand through the use of the DNA polymerase reaction with a small concentration of dideoxynucleoside triphosphates (ddNTPs). These ddNTPs attach to the elongating chain but cause a premature termination due to the lack of OH group in the 3' carbon. Subsequent electrophoresis would give the sequence provided by the differently sized fragments produced in the reactions.

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