title: Enzymatic Method for the Synthesis of Long DNA Sequences with Multiple Repeat Units.
abstract: A polymerase chain reaction (PCR) derived method for preparing long DNA, consisting of multiple repeat units of one to ten base pairs, is described. Two seeding oligodeoxynucleotides, so-called oligoseeds, which encode the repeat unit and produce a duplex with 5'-overhangs, are extended using a thermostable archaeal DNA polymerase. Multiple rounds of heat-cool extension cycles, akin to PCR, rapidly elongate the oligoseed. Twenty cycles produced long DNA with uniformly repeating sequences to over 20 kilobases (kb) in length. The polynucleotides prepared include [A]n /[T]n , [AG]n /[TC]n , [A2 G]n /[T2 C]n , [A3 G]n /[T3 C]n , [A4 G]n /[T4 C]n , [A9 G]n /[T9 C]n , [GATC]n /[CTAG]n , and [ACTGATCAGC]n /[TGACTAGTCG]n , indicating that the method is extremely flexible with regard to the repeat length and base sequence of the initial oligoseeds. DNA of this length (20 kb≈7 μm) with strictly defined base reiterations should find use in nanomaterial applications.
authors.affiliation: Institute for Cell and Molecular Biosciences, University of Newcastle upon Tyne, Newcastle upon Tyne, NE2 4HH (UK).
authors.forename: Colette J
authors.initials: CJ
authors.lastname: Whitfield
authors.affiliation: School of Chemistry, University of Newcastle upon Tyne, Newcastle upon Tyne, NE1 7RU (UK).
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Turley
authors.affiliation: School of Chemistry, University of Newcastle upon Tyne, Newcastle upon Tyne, NE1 7RU (UK).
authors.forename: Eimer M
authors.initials: EM
authors.lastname: Tuite
authors.affiliation: Institute for Cell and Molecular Biosciences, University of Newcastle upon Tyne, Newcastle upon Tyne, NE2 4HH (UK).
authors.forename: Bernard A
authors.initials: BA
authors.lastname: Connolly
authors.affiliation: School of Chemistry, University of Newcastle upon Tyne, Newcastle upon Tyne, NE1 7RU (UK). andrew.pike@ncl.ac.uk.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Pike
mesh_terms: D004247:DNA / Q000138:chemical synthesis* / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004247:DNA
keywords: DNA; DNA structures; nanotechnology; polymerase chain reaction