BMC Genomics BioMed Central Methodology article Open Access Temperature Switch PCR (TSP): Robust assay design for reliable amplification and genotyping of SNPs Tania Tabone1,2, Diane E Mather1 and Matthew J Hayden*1,3 Address: 1Molecular Plant Breeding Co-operative Research Centre and School of Agriculture, Food and Wine, The University of Adelaide, Glen Osmond, SA 5064, Australia, 2Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, VIC 3050, Australia and 3Department of Primary Industries Victoria, Victorian AgriBiosciences Centre, La Trobe R&D Park, Bundoora, VIC 3083, Australia Email: Tania Tabone -
[email protected]; Diane E Mather -
[email protected]; Matthew J Hayden* -
[email protected] * Corresponding author Published: 3 December 2009 Received: 14 August 2009 Accepted: 3 December 2009 BMC Genomics 2009, 10:580 doi:10.1186/1471-2164-10-580 This article is available from: http://www.biomedcentral.com/1471-2164/10/580 © 2009 Tabone et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: Many research and diagnostic applications rely upon the assay of individual single nucleotide polymorphisms (SNPs). Thus, methods to improve the speed and efficiency for single- marker SNP genotyping are highly desirable. Here, we describe the method of temperature-switch PCR (TSP), a biphasic four-primer PCR system with a universal primer design that permits amplification of the target locus in the first phase of thermal cycling before switching to the detection of the alleles.