Increase of Genomic DNA with Verification by SSR Analysis in Carp Carassius auratus

PurposeThe purpose of the present study is the development and optimization of a method for genomic DNA increase in carp Carassius auratus.

MethodThe first step for genomic DNA increase was digestion for genomic DNA by endonuclease. Subsequently, the DNA fragments were linked with specific oligonucleotide adaptors. The DNA fragments were amplified by PCR using the primers that were designed from the sequences of endonuclease recognition sites and adaptors. Finally, the genomic DNA increased.

ResultThe genomic DNA was digested by Taq I endonuclease. Those fragments were linked with specific oligonucleotide adapotors and were amplified by PCR method. The length of PCR product ranged from 250 bp to 1 500 bp for the major DNA fragments. The genomic DNA of carp C. auratus increased by the developed method and was used for microsatellite loci (SSR) amplification. The expected PCR product was obtained for the SSR loci. Compared to the control (genomic DNA was used for PCR), their electrophoresis patterns were the same.

ConclusionThe present study developed a method for genomic DNA increase by PCR in carp C. auratus, which can be applied for SSR analysis and provides the technology support for the massive genetic analysis in fish species.