The University of Southampton
University of Southampton Institutional Repository

Novel approaches in genetic analysis

Novel approaches in genetic analysis
Novel approaches in genetic analysis

The potential use of single nucleotide polymorphisms (SNPs) for genetic mapping and medical diagnostics would require reliable and economical methods for high-throughput SNP genotyping, as the methods in use today are laborious and expensive.

Scorpion primers (PCR primers with a fluorescent probe region attached to the 5’ end) provide a new method for detection of PCR products.  Their unimolecular mechanism of probing brings substantial advantages over other common fluorescent PCR detection methods, especially when the Roche LightCyclerTM thermocycler is employed.  Scorpions have been optimised for PCR detection tests on ABCC7 loci (cystic fibrosis), showing reliability for the discrimination of single nucleotide polymorphisms.

Novel intramolecular TaqMan® ‘intrataq’ probes have also been designed and used in PCR reactions involving SNP discrimination in locus W1282X of the ABCC7 gene.  The ‘intrataq’ probes have been shown to perform better than their intermolecular counterparts in PCR reactions carried out on the Roche LightCyclerTM thermocycler.

The possibility of development of a high-throughput SNP genotyping method using libraries of generic short probes has also been investigated.

University of Southampton
Solinas, Antonio
a3843e95-3cf0-499e-b264-38a2a8ba91eb
Solinas, Antonio
a3843e95-3cf0-499e-b264-38a2a8ba91eb

Solinas, Antonio (2003) Novel approaches in genetic analysis. University of Southampton, Doctoral Thesis.

Record type: Thesis (Doctoral)

Abstract

The potential use of single nucleotide polymorphisms (SNPs) for genetic mapping and medical diagnostics would require reliable and economical methods for high-throughput SNP genotyping, as the methods in use today are laborious and expensive.

Scorpion primers (PCR primers with a fluorescent probe region attached to the 5’ end) provide a new method for detection of PCR products.  Their unimolecular mechanism of probing brings substantial advantages over other common fluorescent PCR detection methods, especially when the Roche LightCyclerTM thermocycler is employed.  Scorpions have been optimised for PCR detection tests on ABCC7 loci (cystic fibrosis), showing reliability for the discrimination of single nucleotide polymorphisms.

Novel intramolecular TaqMan® ‘intrataq’ probes have also been designed and used in PCR reactions involving SNP discrimination in locus W1282X of the ABCC7 gene.  The ‘intrataq’ probes have been shown to perform better than their intermolecular counterparts in PCR reactions carried out on the Roche LightCyclerTM thermocycler.

The possibility of development of a high-throughput SNP genotyping method using libraries of generic short probes has also been investigated.

Text
905433.pdf - Version of Record
Available under License University of Southampton Thesis Licence.
Download (39MB)

More information

Published date: 2003

Identifiers

Local EPrints ID: 465017
URI: http://eprints.soton.ac.uk/id/eprint/465017
PURE UUID: 4bfda91f-2de6-4095-827a-6f245a72521e

Catalogue record

Date deposited: 05 Jul 2022 00:16
Last modified: 16 Mar 2024 19:53

Export record

Contributors

Author: Antonio Solinas

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

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×