The SNaPshot® assay is commonly used for forensic SNP analysis. However, it is a multi-step process with potential post-PCR contamination risk. The single tube high resolution melting (HRM) temperature real-time PCR method is an alternative, eliminating the post-PCR tube transfer of SNaPshot®. Eight individual DNA samples were genotyped at the six IrisPlex SNP loci using both the IrisPlex published primer set and a set of custom designed HRM primers. The performance of MeltDoctor™ (Life Technologies®) and SensiFast™ (Bioline®) HRM mastermixes was examined on the ViiA™ 7 Real Time PCR platform for 10 ng and 1 ng DNA template amounts. The resultant genotypes were compared with those derived from SNaPshot®. This preliminary study demonstrates HRM potentially offers a fast and flexible alternative to SNaPshot® for small numbers of SNP loci without the associated contamination risk from post-PCR processes.
|Number of pages
|Forensic Science International: Genetics Supplement Series
|Published - 2013