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Biology, 21.03.2020 10:28 cbbentonam72

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Suppose you have a PCR amplicon consisting of the sequence below. The primers used to generate the amplicon are: 5'AT3' and 5'GA3'. You can use one of these primer sequences for a subsequent sequencing reaction but not both simultaneously because they will generate sequence information for different nucleotide chains. The chains are complementary but not identical therefore fragments of the same size will be likely to contain different chain terminating nucleotides. For example, 4 base long sequence fragments using 5'AT3' will terminate in a G, whereas 4 base long sequence fragments using 5'GA3' will terminate in a T.

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