Genomics and Applied Biology 2018, Vol.9, No.4, 19-23
22
gelatin), 0.1 mmol/LdNTP, 0.2 μmol/L primer concentration, 1.0 U
Taq
enzyme (Biostar International). Executed
SSR reaction program: pre-denatured 4 min at 94°C, then denatured 30 s per cycle at 94°C, annealed 30 s, and
extended 30 s at 72℃. After 35 cycles, the final extension step was carried out at 72℃ for 10 min (Xie et al.,
2007). AFLP experiment referred to Vos (1995), in which
Eco
RⅠ (6 recognition base sites GAATCC) and
Mse
Ⅰ (4
recognition base sites TTAA) were selected as the restriction endonuclease. Both endonuclease and ligase bought
from New England Biolab (Xie et al., 2011).
3.2.3 Data statistics and analysis
The "0-1" identification and labeling system was used to observe whether the amplified bands were labeled in the
gel electrophoresis atlas obtained from the amplified products of DNA fragments. If there were amplified bands,
marked "1", if there were no amplified bands, marked "0", recorded the location of the labeled bands as well.
POPGENE software (Yeh et al., 1999,
) was used to calculate average
genetic diversity, according to the method of Nei (1973). Genetic identity and genetic distance among populations
before and after the correction of the deviation were calculated, according to the methods of Nei (1972) and Nei
(1978). ARLEQUIN software (Schneider et al., 2000,
) was
used to calculate the
Fst,
according to the method of Weir and Cockerham (1984) (Xie et al., 2007; 2011). The
ratio of polymorphic loci, population specific
Fst
indices, average genetic diversity, genetic identity, and genetic
distance among populations were calculated by SSR and AFLP respectively.
Autho
r
s’ contributions
XXY was the executor of
this study who completed the design and study of experiment, carried out data statistics and analysis, and
drafted and revised the manuscript. He's also the head of one of the funding projects. ZJX participated in the manuscript revision.
LSF was the designer and director of another funding project. All authors read and approved the final manuscript.
Acknowledgments
This study was supported by a grant from the National Key Technologies R & D Program of China during the 10th Five-Year Plan
Period (2001BA505B0513) and the Central Level, Scientific Research Institutes for Basic R & D Business Special Fund (South
China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences) (2007TS04). Thanks to Shanghai Ocean University
for providing Nile tilapia materials for this study.
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