A C G T An unknown DNA template was sequenced using Sanger\'s chain termination

Question

A C G T An unknown DNA template was sequenced using Sanger's chain termination method and separated by agarose gel electrophoresis on 20mm gel length. The grid is equally divided on the agarose gel and each grid represents 1bp (base pair) size of DNA. The primer that was used in the sequencing reaction is: 3'-AAGGTTCCG-5' (a) How many fragments were produced during the sequencing reaction? (b) Determine the template sequence of the shortest and longest fragment on the agarose gel (c) What is the size of the DNA template including the primer? (d) Read the unknown template sequence on the agarose gel and label the ends (e) Does the template sequence contain any gene? If yes, determine the mRNA transcript and the a mino acid chain produced (f) Assuming the unknown sequence contains a gene expressed by a tissue and you have three minute mRNA samples extracted from three different tissues labeled A, B and C. If the amount of mRNA is not sufficient for blotting, describe how you can identify the tissue that is expressing the gene (g) Describe how you can use gene micro array to determine if an unknown DNA sample contains a gene that causes a known disease

Explanation / Answer

Primer sequence: 5'-GCCTTGGAA-3'

A. There are 21 bands on the gel. So, 21 products are produced.

B. Shortest template sequence: 3'-CGGAACCTTG-5'

Longest template sequence: 3'-CGGAACCTTGGTTACGAGCAGTGAATCTTC-5'

C. Size of the template including the primer = 9 + 21 = 30 ntd

D. Template strand sequence = 3'-CGGAACCTTGGTTACGAGCAGTGAATCTTC-5'

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