Basic HTML Version
International Journal of Aquaculture 2012, Vol.2, No.2, 5
-
10
http://ija.sophiapublisher.com
9
City, CA). PCR was conducted on a Master Cycler
Gradient (Eppendorf, Germany) with the following
steps: 94
℃
for 4 min followed by 34 cycles of 94
℃
for 30 sec, 30 sec annealing (temperatures indicated in
Table 1) for 45 s, followed by 10 cycles of 94
℃
for
30 sec, 53
℃
for 45 sec, 72
℃
for 45 sec, and a final
extension at 72
℃
for 10 min. Amplified products
were detected and sized on an ABI 3130 Prism
Genetic Analyzer (Applied Biosystems) using
GS-500LIZ (Applied Biosystems) as the size standard.
Allele scoring was performed with GeneMapper v3.5
(Applied Biosystems). Samples failing to amplify at
the first time were re-amplified once.
3.4 Statistical analysis
The Micro-Checker 2.2.1 software (van Oosterhout et
al., 2004) was used to identify possible genotyping
errors and null alleles (1 000 randomizations).
GENEPOP on the Web (http://genepop.curtin.edu.au/)
was used to test deviations from Hardy-Weinberg
equilibrium (HWE) for each locus and linkage
disequilibrium between all pairs (exact tests, 1 000
iterations). ARLEQUIN 3.0 software (Excoffier et al.,
2005) was used to calculate the observed (
H
O
) and
expected (
H
E
) heterozygosity. All tests were corrected
for multiple comparisons by Bonferroni’s correction
(Rice, 1989).
Author Contributions
NZ and YQ are the executor of experimental research in this
study, they have the same contribution to the paper. YW and
XZH make the experimental design, data analysis, paper
writing and revising. AMW is the supervisor of the project and
XFC provides the necessary facilities for the research to be
performed.
Acknowledgement
This study was supported by the National Natural Science
Foundation of China (40866003 and 31060354), the Natural
Science Foundation of Hainan (80616, 311024), and the
National Basic Research Program of China (973 program,
2010CB126405).
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