Oligonucleotide and Long Polymeric DNA Encoding

Miller, E ; Mariella Jr., R P ; Christian, A T ; Gardner, S N ; Williams, J M

Lawrence Livermore National Laboratory

关键词: Thermodynamics;    Dna;    Genes;    Microelectronics;    Functionals;   

DOI  :  10.2172/15009752
RP-ID  :  UCRL-TR-201189
RP-ID  :  W-7405-ENG-48
RP-ID  :  15009752

美国|英语

来源: UNT Digital Library

【 摘 要 】

This report summarizes the work done at Lawrence Livermore National Laboratory for the Oligonucleotide and Long Polymeric DNA Encoding project, part of the Microelectronic Bioprocesses Program at DARPA. The goal of the project was to develop a process by which long (circa 10,000 base-pair) synthetic DNA molecules could be synthesized in a timely and economic manner. During construction of the long molecule, errors in DNA sequence occur during hybridization and/or the subsequent enzymatic process. The work done on this project has resulted in a novel synthesis scheme that we call the parallel pyramid synthesis protocol, the development of a suit of computational tools to minimize and quantify errors in the synthesized DNA sequence, and experimental proof of this technique. The modeling consists of three interrelated modules: the bioinformatics code which determines the specifics of parallel pyramid synthesis for a given chain of long DNA, the thermodynamics code which tracks the products of DNA hybridization and polymerase extension during the later steps in the process, and the kinetics model which examines the temporal and spatial processes during one thermocycle. Most importantly, we conducted the first successful syntheses of a gene using small starting oligomers (tetramers). The synthesized sequence, 813 base pairs long, contained a 725 base pair gene, modified green fluorescent protein (mGFP), which has been shown to be a functional gene by cloning into cells and observing its green fluorescent product.