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1. A Sanger dideoxy approach was used to determine the sequence of a gene X. Res

ID: 221229 • Letter: 1

Question

1. A Sanger dideoxy approach was used to determine the sequence of a gene X. Results of the DNA sequencing were shown below on the gel. Samples were loaded on the top of the gel. The positive electrode is located at the bottom of the gel. What is the sequence of the template DNA that was used to generate the Sanger dideoxy sequencing gel shown below? Be sure to indicate the 5' and 3 ends. Note that the letter above each lane indicates the dideoxynucleotide that was present in the tube whose reaction was loaded into that lane. (12 points)

Explanation / Answer

The sequence of the read sequence is 5' TGAACGCATGCT 3'. ( The smallest fragment indicating the 5' end of the newly synthesisized strand; Since the smallest band will travel fastest and hence travel the maximum distance, as per the figure this nucleotide is be T). The template strand will be having a sequence complementary to this.

Read sequence   5' TGAACGCATGCT 3'

Template sequence 3' ACTTGCGTACGA 5'