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sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE)
and isoelectric focusing (IEF) (Huang, Jeng, Chen, & Hwang,
2008). However, compared with proteins, the DNA extracted from
processed products is more suitable for species identification
because it is more stable and is less affected by thermal treatment
(Pardo & Perez-Villareal, 2004).
DNA-based methods offer an alternative approach to species
identification by using polymerase chain reaction (PCR). These
methods include the following: nested PCR (Pardo & Perez-
Villareal, 2004; Zhang, Huang, Cai, & Huang, 2007), single nucleotide
polymorphism (SNP) analysis (Itoi et al., 2005), multiplex PCR
(Trotta et al., 2005), real-time PCR (Itoi et al., 2005; Trotta et al.,
2005), PCR-restriction fragment length polymorphism (PCR-RFLP)
analysis (Chen, Hsieh, & Hwang, 2012; Hsieh et al., 2010), PCRsingle
strand conformation polymorphism (PCR-SSCP) analysis
(Rehbein, Kress, & Schmidt, 1997), denaturing gradient gel electrophoresis
(DGGE) (Comi et al., 2005), and forensically informative
nucleotide sequencing (FINS) (Chen et al., 2012). Many previous
works indicated that the direct sequencing of PCR-amplified
products using PCR-RFLP or PCR-SSCP can be used to identify the
species of heavily processed products, such as canned sardines
(Infante, Catanese, Ponce, & Manchado, 2004).
Although PCR-RFLP is useful for species detection, this method
depends on using gel electrophoresis and staining with chemical
reagents that are potentially hazardous, and it is time-consuming
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