How Does Sanger Sequencing Work?
What are the steps involved in Sanger sequencing?
Choose the correct sequence from the options below:
A. Denaturation, Primer annealing, Extension
B. Extension, Denaturation, Primer annealing
C. Primer annealing, Extension, Denaturation
D. Denaturation, Extension, Primer annealing
Answer:
The steps of Sanger sequencing are denaturation, primer annealing, and extension.
Sanger sequencing is a method used to determine the sequence of nucleotides in a DNA molecule. It involves the use of DNA polymerase, primers, and dideoxynucleotides (ddNTPs) labeled with fluorescent markers.
The steps of Sanger sequencing are as follows:
Denaturation:The DNA sample is heated to high temperatures to separate it into two strands.
Primer annealing:Short DNA segments called primers are added, which have sequence complementary to the target DNA.
Extension:DNA polymerase and all four nucleotides (A, T, G, and C) are added, allowing the synthesis of the new DNA strand.
The sequence is read using a laser scanner that detects the fluorescent markers on the dideoxynucleotides, which terminate the synthesis of the DNA strand at specific points.