Andrew Pike

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Organization: Newcastle University , England
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Title: Lecture(PhD)
Co-reporter:Colette J. Whitfield;Andrew T. Turley;Dr. Eimer M. Tuite; Bernard A. Connolly;Dr. Andrew R. Pike
Angewandte Chemie 2015 Volume 127( Issue 31) pp:9099-9102
Publication Date(Web):
DOI:10.1002/ange.201502971

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.

Co-reporter:Colette J. Whitfield;Andrew T. Turley;Dr. Eimer M. Tuite; Bernard A. Connolly;Dr. Andrew R. Pike
Angewandte Chemie International Edition 2015 Volume 54( Issue 31) pp:8971-8974
Publication Date(Web):
DOI:10.1002/anie.201502971

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.

Co-reporter:Shahrbanou MoradpourHafshejani, Joseph H. Hedley, Alexandra O. Haigh, Andrew R. Pike and Eimer M. Tuite  
RSC Advances 2013 vol. 3(Issue 39) pp:18164-18172
Publication Date(Web):12 Jul 2013
DOI:10.1039/C3RA43090A
Proflavine diazide (PD) with amido-azide substituents on the amine groups and its N-methylated analogue (MePD) bind strongly to DNA by nearest-neighbour intercalation with little sequence selectivity, presenting reactive azide groups in the major groove. PD is neutral in aqueous solution but experiences binding-coupled protonation on interaction with DNA with an apparent pKa shift of 2.5 units. MePD can be click modified in situ on DNA with alkyne-functionalised thienyl-pyrrole as a precursor for conducting polymer synthesis, and remains intercalated after reaction with the substituents aligned in the groove.
CYTIDINE, 5'-O-[BIS(4-METHOXYPHENYL)PHENYLMETHYL]-2'-DEOXY-5-IODO-
1H-PYRROLE, 1-(10-UNDECENYL)-
ACETAMIDE, N,N'-3,6-ACRIDINEDIYLBIS[2-CHLORO-
Ethanamine, 2-azido-
1-Propanamine, 3-azido-
1H-Pyrrole, 1-(2-propenyl)-
1H-Pyrrole,1-(4-pentynyl)-(9CI)
Thiophene, 2-(2-pyrrolyl)-
1H-Pyrrole, 1-(2-propynyl)-
hexammineruthenium