Dideoxynucleotide triphosphates (ddNTPs) are used in manual Sanger sequencing and in many automated methods of sequencing. What is the purpose of using dideoxynucleotides in DNA sequencing? Select ALL that apply. The figure below may be helpful.Multiple choice

Nucleotides are added at the 3' hydroxyl group (–OH) of the growing chain during DNA synthesis.
ddNTPs lack a free hydroxyl (–OH) group at the 2' carbon, thus preventing additional nucleotides from being added to the chain when a ddNTP is incorporated.
ddNTPs lack a free hydroxyl (–OH) group at the 3' carbon, thus preventing additional nucleotides from being added to the chain when a ddNTP is incorporated.
Nucleotides are added at the 5' hydroxyl group (–OH) of the growing chain during DNA synthesis.
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A primer used in dideoxy DNA sequencing is 20 nucleotides long and has the following sequence: 5’-GGATCCATGACTAGTCCGAC-3’. A segment of DNA is cloned into a vector and then the vector DNA is denatured and subjected to dideoxy DNA sequencing method. Below is the DNA sequence from a region of the vector. The primer-annealing site is shown in bold and underlined. 3-CCCGATCGGCCTAGGTACTGATCAGGCTGAATGACT-5’ Based on the sequence above, what would be the size(s) of the band(s) (i.e., the number of nucleotides in each band) in which dideoxyT had been added to the sequencing reaction?
2. How do dideoxynucleotide triphosphates (diNTPs) in the Sanger method facilitate DNA sequencing?
ddATP, ddTTP, ddCTP, and ddGTP are labeled with red, green, yellow, and blue fluorescent dyes, respectively during Sanger sequencing. A five-base read from a sequencing reaction produced the following color sequence, read by the computer: red, yellow, yellow, green, green. What is the sequence of the template DNA?
Question19 How is the DNA sequence determined in Automated Sanger sequencing after the chain termination reactions are complete? Select one alternative: By separating fluorescently labelled fragments by size in a capillary gel and reading the colour of the terminal base. By detecting the change in concentration as each nucleotide is incorporated. By generating clusters on a flow cell and imaging the incorporation of reversible terminators. Bypassing millions of DNA strands through a nanopore and measuring electrical current changes. ResetMaximum marks: 1 Flag question undefined
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