Question

When setting up a Sanger sequencing reaction, each reaction should include template DNA, nucleotides, dideoxynucleotides, buffer, DNA polymerase, and _____.
a. Forward and reverse primers
b. Forward or reverse primers
c. Forward and reverse probes
d. Forward or reverse probes

Answer 1

Answer:

b. Forward or reverse primers

Explanation:

Sanger sequencing is a technique of DNA sequencing based on the extension of DNA fragments with variable sizes terminated with dideoxynucleotides at the 3′ end. This technique was developed by Frederick Sanger in 1977. In Sanger sequencing, a short primer is added in order to bind by complementarity to the target DNA region of interest. Subsequently, a DNA polymerase adds nucleotides (A, T, C and G) in the 5'-3' direction. Finally, the extension of the DNA strand is stopped by adding dideoxynucleotides, which are nucleotide analogs (i.e., modified nucleotides) that act as DNA synthesis terminators.