labeo, Pseudorasbora parva, Rhodeus ocellatus ocellatus, Varicorhinus
barbatulus, and Zacco pachycephalus. Based on their morphological
characteristics, these species of fish were identified by biologist Dr.
Hong-Ming Chen. Twenty-four samples of Cyprinidae products
were collected from traditional markets in Taipei (T1eT10) and
from internet shops (I1eI14) in Taiwan. All of the samples were
transferred immediately to the laboratory on ice.
2.2. Design of primers
To obtain the complete mitochondrial cytb gene sequence of
each of the 10 Cyprinidae species, we used 2 sets of primers to
amplify the cytb fragments: (1) L14734-Glu (50-AAC CAC CGT TGT
TAT TCA ACT-30) and H15557-CYB (50-GGC AAA TAG GAA RTA TCA
YTC-30); and (2) Cytb-F (50-CAC GAR ACR GGR TCN AAY AA-30) and
Thr-R (50-ACC TCC RAT CTY CGG ATT ACA-30) (Chen et al., 2012). For
the RFLP analysis, we designed a new primer set for CypbL (50-AAC
CAC ATA ATG GCA AGC CT-30) and CypbH (50-GGT ADG ATC CRT ART
ADA GGC C-30), which was used to amplify a 331-bp partial cytb
fragment of Cyprinidae and was further analyzed in this study.