DNA Sequencing The following DNA fragment was sequenced by the Sanger method. A
ID: 81260 • Letter: D
Question
DNA Sequencing The following DNA fragment was sequenced by the Sanger method. A sample of the DNA was reacted with DNA polymerase and each of the nucleotide mixtures listed below. Dideoxynucleotides (ddNTPs) were added in relatively small amounts. dATP, dTTP, dCTP, dGTP, ddTTP dATP, dTTP, dCTP, dGTP, ddGTP dATP, dCTP, dGTP, ddTTP dATP, dTTP, dCTP, dGTP, ddCTP dATP, dTTP, dCTP, dGTP The resulting DNA was separated by electrophoresis on an agarose gel, and the fluorescent bands on the gel were located. Assuming that all mixtures were run on the same gel, what did the lanes of the gel look like? Write how many band/relative position from each of the five reactions.Explanation / Answer
Lane 1: The response mixture that produce these bands incorporate all the dideoxythymidine + deoxynucleotides. The particle are of different distance, all terminating. Where a ddTTP was replacement for a dTTP. For a little segment of the strands synthesized in the research, ddTTP would not be placed and the sequence would thus expand to the end of G. Thus, the 9 sequence are:
TAATGCGTTCCTGTAATCTG - 5-primer
TAATGCGTTCCTGTAATCT - 5-primer
TAATGCGTTCCTGTAAT - 5-primer
TAATGCGTTCCTGT - 5-primer
TAATGCGTTCCT - 5-primer
TAATGCGTT- 5-primer
TAATGCGT - 5-primer
TAAT - 5-primer
5-primer-T
Lane 2: Identically , this track will have 4 stripe (apex to base), for the following particle, each ending where ddGTP was placed in location of dGTP:
TAATGCGTTCCTGTAATCTG - 5-primer
TAATGCGTTCCTG - 5-primer
TAATGCG - 5-primer
TAATG - 5-primer
Lane 3: Because combination 3 deficiency dTTP, every particle was ended instantly after the primer as ddTTP was placed to construct 5-primer-T. The output will be a alne thick band close to the base of the gel.
Lane 4: The deoxynucleotides were assumption, but no dideoxynucleotide, a lone tag product constructed:
TAATGCGTTCCTGTAATCTG - 5-primer
This will become visible as a alone thick stripe at the apex of the gel.
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