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- Title
Enzymatic Method for the Synthesis of Long DNA Sequences with Multiple Repeat Units.
- Authors
Whitfield, Colette J.; Turley, Andrew T.; Tuite, Eimer M.; Connolly, Bernard A.; Pike, Andrew R.
- 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, [A2G] n/[T2C] n, [A3G] n/[T3C] n, [A4G] n/[T4C] n, [A9G] n/[T9C] 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.
- Subjects
NUCLEOTIDE sequence; ENZYMATIC analysis; POLYMERASE chain reaction; DNA polymerases; NANOSTRUCTURED materials
- Publication
Angewandte Chemie, 2015, Vol 127, Issue 31, p9099
- ISSN
0044-8249
- Publication type
Article
- DOI
10.1002/ange.201502971