Non-gel based enzyme assays and combinatorial screening of linker-dye combinations
Non-gel based enzyme assays and combinatorial screening of linker-dye combinations
An understanding of the function and role of nuclei acids has grown rapidly since the structure of DNA was first postulated in 1953. Modified nucleosides and nucelotides now play an important role both as therapeutic agents and as probes for obtaining information concerning the structure and sequence of DNA. This project set out to develop a new method of evaluating target compounds designed for DNA sequencing. Combinatorial chemistry and solid phase synthesis have revolutionised the process of drug discovery since their introduction but have not been widely applied in nucleoside chemistry. Realising the value of modified nucleosides and the advantages of solid phase chemistry it was an attractive proposition to combine the two to allow the preparation of libraries of nucleoside analogues. A thymidine-based resin was therefore developed and successfully used for the synthesis of libraries of nucleoside analogues. The resin proved to be well suited for SPPS when libraries of compounds were easily synthesised using this resin.
A novel DNA sequencing-by-synthesis strategy has been proposed in which single stranded DNA fragments are immobilised on a solid support. The primer is extended one base at a time using nucleotides that carry a reporter moiety attached through a peptide spacer. To optimise the peptide functionality for cleavage and incorporation a library was synthesised on the thymidine resin and screened for efficient substrates. Several good substrates were identified using this approach. To further improve the throughput of the method a biocompatible thymidine resin was also prepared on which a smaller library was synthesised. On-bead screening proved successful and target sequences could rapidly be identified.
University of Southampton
Johnson, Karin
4f5d1776-37a7-47c4-8e7e-40311fefce91
2003
Johnson, Karin
4f5d1776-37a7-47c4-8e7e-40311fefce91
Johnson, Karin
(2003)
Non-gel based enzyme assays and combinatorial screening of linker-dye combinations.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
An understanding of the function and role of nuclei acids has grown rapidly since the structure of DNA was first postulated in 1953. Modified nucleosides and nucelotides now play an important role both as therapeutic agents and as probes for obtaining information concerning the structure and sequence of DNA. This project set out to develop a new method of evaluating target compounds designed for DNA sequencing. Combinatorial chemistry and solid phase synthesis have revolutionised the process of drug discovery since their introduction but have not been widely applied in nucleoside chemistry. Realising the value of modified nucleosides and the advantages of solid phase chemistry it was an attractive proposition to combine the two to allow the preparation of libraries of nucleoside analogues. A thymidine-based resin was therefore developed and successfully used for the synthesis of libraries of nucleoside analogues. The resin proved to be well suited for SPPS when libraries of compounds were easily synthesised using this resin.
A novel DNA sequencing-by-synthesis strategy has been proposed in which single stranded DNA fragments are immobilised on a solid support. The primer is extended one base at a time using nucleotides that carry a reporter moiety attached through a peptide spacer. To optimise the peptide functionality for cleavage and incorporation a library was synthesised on the thymidine resin and screened for efficient substrates. Several good substrates were identified using this approach. To further improve the throughput of the method a biocompatible thymidine resin was also prepared on which a smaller library was synthesised. On-bead screening proved successful and target sequences could rapidly be identified.
Text
914701.pdf
- Version of Record
More information
Published date: 2003
Identifiers
Local EPrints ID: 465117
URI: http://eprints.soton.ac.uk/id/eprint/465117
PURE UUID: aba924eb-8b59-4eba-b59e-b962987d75fa
Catalogue record
Date deposited: 05 Jul 2022 00:24
Last modified: 16 Mar 2024 19:57
Export record
Contributors
Author:
Karin Johnson
Download statistics
Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.
View more statistics