Fredrick Sanger invented the first method for sequencing DNA, which took advanta

Question

Fredrick Sanger invented the first method for sequencing DNA, which took advantage of knowing the mechanism of DNA Polymerase. His method utilized 2',3'-dideoxynucleotide triphosphates as chain-terminating substrate analogs.

A.) What substrate analogs could be used to sequence RNA using RNA polymerase?_________________________

B.) A DNA was sequenced using the original Sanger method. The sequencing reactions were analyzed by gel electrophoresis (shown below). Eachreaction contained a different substrate analog (labeled in the gel lanes as A, G, C, and T). A primer was paired with the template strand of the DNA. Marked on left side of this gel is gel band of radioactively labeled primer. In the space provided below, complete the base sequence of the template DNA stranded.

Template strand: 3'- CGAAGGAG

Primer: 5'- GCTTCCTC

Explanation / Answer

Nucleotide triphosphate substrate analogues; Poly T-oligos are widely acts as substrate analogues to sequence RNA. 5-(3-Aminopropyl) uridine triphosphate, analogues of 2-deoxy-2-fluoro- and 2-deoxy-2-amino compounds are widely used for RNA sequencing.

Template: 3’-CGAAGGAG GCATCGCTAATAGC-5’

Primer:    5’-GCTTCCTCCGTAGCGATTATCG-3’

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