High pressure liquid chromatography of oligodeoxyribonucleotides

Cook, Alan F.; Czekala, Alice De; Gabriel, Thomas F.; Harvey, Clifford; Holman, Michael J.; Michaelewsky, J.E.; Nussbaum, A. L.

doi:10.1016/0005-2787(73)90288-8

Cited by 11 publications

(1 citation statement)

References 14 publications

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“…The synthesis of a DNA duplex coding for an S-peptide analog of bovine pancreatic ribonuclease A (1) led to the study of its enzymatic multiplication. There is need for a method to increase the original quantity of a chemically synthesized DNA sequence after its laborious assembly.…”

Section: Introductionmentioning

confidence: 99%

Enzymatic multiplication of a chemically synthesized DNA fragment

Olson¹,

Gabriel²,

Michalewsky³

et al. 1975

Nucl Acids Res

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A synthetic DNA fragment of 19 residues was enlarged by the enzymatic addition of deoxyadenylate residues to its 3'-end with calf thymus terminal deoxynucleotidyl transferase. The 3'-terminus of this elongated DNA strand was blocked with 2',3'-dideoxyadenylate to prevent hydrolysis by the 3'-exonuclease function of E. coli DNA polymerase I. This elongated and 3'-blocked fragment was annealed to an oligomeric primer and used as a template for the synthesis of a complementary copy of the synthetic 19-mer. The product of such a repair synthesis was separated by gel filtration and analyzed by nearest neighbor techniques.All template strands were copied with complgte repair in over 90% of the chains. Facile recovery of the elongated template by virtue of its size permitted repetition of the copy process, thus allowing accumulation of the desired strand.

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Section: Introductionmentioning

confidence: 99%