Question

In the original Sanger sequencing method, you obtain a readout that has 4 lanes, each lane corresponding to a nucleotide. If you get the results back and only 3 of the 4 lanes show the expected bands, which of the following would NOT be a reason for your results?a. The fluorescent color used for the label was not the correct color.b. The sample was not loaded in the well for the blank lane.c. No dNTPs were added for the blank lane's reaction.d. No sequencing primer was added for the blank lane's reaction.e. No ddNTP was added for the blank lane's reaction.

Answer 1

Answer:

Option-1

Explanation:

DNA sequencing refers to the method which analyses the DNA and determines the sequence of nucleotide.

The Sanger method uses the same things used during the DNA replication process but an additional thing is also there called the dideoxynucleotides attached with fluorescent dye.

When thus ddNTP binds to the growing chain, then the replication process stops as ddNTP lacks the -OH group at 3'position.In the given question, since only 3 bands shows the result but one band does not it means that the florescent label has not been recognized as ddNTP label.

Thus, the problem lies in the florescent label and therefore option-1 is correct.