Determination of the 3' terminal nucleotide of DNA fragments

AUTOR(ES)

Olson, Kenneth

RESUMO

A new method for determining the 3′-terminal nucleotide of a DNA strand is presented. Use is made of the fact that one (and only one) 2′,3′-dideoxyribonucleotide can be added to the 3′-end of a DNA fragment with calf thymus terminal transferase. Addition of more than one nucleotide analog per strand is impossible due to the absence of a 3′terminal hydroxyl group. If the terminating dideoxyribonucleotide contains an [α32P] label, the resulting 3′-blocked strand can be digested by “nearest neighbor” techniques and the original 3′-endgroup determined. Picomole quantities of DNA strands can be labeled and the 3′end determined.

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