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.