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(4 marks) A fragment of DNA is cloned into a plasmid with a sequencing primer bi

ID: 301459 • Letter: #

Question

(4 marks) A fragment of DNA is cloned into a plasmid with a sequencing primer binding site. After dideoxy sequencing, the gel pattern shown in this diagram is obtained ddATP reaction ddTTP reaction ddCTP reaction ddGTP reaction (a) What was the sequence of the complementary DNA strand in the sequencing reaction? 5 3' (b) What was the sequence of the template DNA in the sequencing reaction?5' 3' Please write your sequencing in the correct 5' to 3' direction. Deductions will be made for sequences input in the incorrect direction.

Explanation / Answer

In dideoxy sequencing, the gel is read from bottom to top in 5’-->3’ direction

5’TGACACGT 3’

(a)Complementary strand in the sequence reaction is 3’ ACTGTGCA 5’

(b)Template for the sequence of template DNA is 3’ ACTGTGCA 5’