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. 2008 Dec;2(3-4):67-73.
doi: 10.1007/s11693-009-9023-x. Epub 2009 Apr 3.

A theoretical model for template-free synthesis of long DNA sequence

S M Minhaz Ud-Dean  1
Affiliations

A theoretical model for template-free synthesis of long DNA sequence

S M Minhaz Ud-Dean. Syst Synth Biol. 2008 Dec.

Abstract

This theoretical scheme is intended to formulate a potential method for high fidelity synthesis of Nucleic Acid molecules towards a few thousand bases using an enzyme system. Terminal Deoxyribonucleotidyl Transferase, which adds a nucleotide to the 3'OH end of a Nucleic Acid molecule, may be used in combination with a controlled method for nucleotide addition and degradation, to synthesize a predefined Nucleic Acid sequence. A pH control system is suggested to regulate the sequential activity switching of different enzymes in the synthetic scheme. Current practice of synthetic biology is cumbersome, expensive and often error prone owing to the dependence on the ligation of short oligonucleotides to fabricate functional genetic parts. The projected scheme is likely to render synthetic genomics appreciably convenient and economic by providing longer DNA molecules to start with.

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Figures

Fig. 1
Fig. 1
Simplified scheme for TdT based DNA synthesis
Fig. 2
Fig. 2
Lack of precise temporal order in removal of excess modified NTP and deacetylation may lead to unintended chain elongation
Fig. 3
Fig. 3
Proposed protocol for DNA synthesis utilizing the scheme
See this image and copyright information in PMC

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