Esox reieherti Dybowsk genomic microsateUites were developed by using enrichment protocols combined with radioactive hybridization protocol. Four hundred to nine hundred base pair fragments were selected for the whole...Esox reieherti Dybowsk genomic microsateUites were developed by using enrichment protocols combined with radioactive hybridization protocol. Four hundred to nine hundred base pair fragments were selected for the whole genome. DNA PCR amplification after digestion with restriction endonuclease Sau 3A Ⅰ, and (CA)12, (GA)12 probes marked with biotin were used for microsateUite DNA enrichment. The product fragments were connected with carder pGEM-T and transferred into DH5α Escherichia coli competent cells, and radioactive isotope probes marked with γ^-32 p were used for the second hybridization. As a result, a total of 1600 bacteria were obtained in the microsatellite genomic libraries, positive clones accounted for 90.91% before hybridization and 81.25% after hybridization, amounting to 1300. One hundred and ninety-six positive clones were selected for sequencing, and 192 clones included microsateUite sequences. The microsateUite sequences obtained, mono-nucleotide, quad-nucleotide and quint-nucleotide repeat motifs were observed beside double-base-pairs CA/GT, GA/CT. Seventy primers were designed according to the flanking sequences by using software Primer Premier 5.0, and 32 primers were selected to be synthesized. After optimizing PCR reaction conditions, 28 primers were amplified and produced clear purpose bands. The aim of our research was to promote the development and utilization of E. reieherti genomic resource, and lay the foundation for optimizing E. reieherti breeding strain in order to detect the genetic diversity and construct a genetic map.展开更多
A novel oxygen-enriched method is presented. Using two opposite magnetic poles of two magnets with certain distance forms a magnetic space having a field intensity gradient near its borders. When air injected into the...A novel oxygen-enriched method is presented. Using two opposite magnetic poles of two magnets with certain distance forms a magnetic space having a field intensity gradient near its borders. When air injected into the magnetic space outflows from the magnetic space via its borders, oxygen molecules in air will experience the interception effect of the gradient magnetic field, but nitrogen molecules will outflow without hindrance. Thereby the continuous oxygen enrichment is realized. The results show that the maximum increment of oxygen concentration reaches 0.49% at 298 K when the maximum product of magnetic flux density and field intensity gradient is 563T^2/m. The enrichment level is significantly influenced by the gas temperature and the magnetic field. The maximum increment of oxygen concentration drops to 0.16% when the gas temperature rises to 343 K, and drops to 0.09% when the maximum product of magnetic flux density and gradient is reduced to 101 T^2/m from 563 T^2/m.展开更多
文摘Esox reieherti Dybowsk genomic microsateUites were developed by using enrichment protocols combined with radioactive hybridization protocol. Four hundred to nine hundred base pair fragments were selected for the whole genome. DNA PCR amplification after digestion with restriction endonuclease Sau 3A Ⅰ, and (CA)12, (GA)12 probes marked with biotin were used for microsateUite DNA enrichment. The product fragments were connected with carder pGEM-T and transferred into DH5α Escherichia coli competent cells, and radioactive isotope probes marked with γ^-32 p were used for the second hybridization. As a result, a total of 1600 bacteria were obtained in the microsatellite genomic libraries, positive clones accounted for 90.91% before hybridization and 81.25% after hybridization, amounting to 1300. One hundred and ninety-six positive clones were selected for sequencing, and 192 clones included microsateUite sequences. The microsateUite sequences obtained, mono-nucleotide, quad-nucleotide and quint-nucleotide repeat motifs were observed beside double-base-pairs CA/GT, GA/CT. Seventy primers were designed according to the flanking sequences by using software Primer Premier 5.0, and 32 primers were selected to be synthesized. After optimizing PCR reaction conditions, 28 primers were amplified and produced clear purpose bands. The aim of our research was to promote the development and utilization of E. reieherti genomic resource, and lay the foundation for optimizing E. reieherti breeding strain in order to detect the genetic diversity and construct a genetic map.
文摘A novel oxygen-enriched method is presented. Using two opposite magnetic poles of two magnets with certain distance forms a magnetic space having a field intensity gradient near its borders. When air injected into the magnetic space outflows from the magnetic space via its borders, oxygen molecules in air will experience the interception effect of the gradient magnetic field, but nitrogen molecules will outflow without hindrance. Thereby the continuous oxygen enrichment is realized. The results show that the maximum increment of oxygen concentration reaches 0.49% at 298 K when the maximum product of magnetic flux density and field intensity gradient is 563T^2/m. The enrichment level is significantly influenced by the gas temperature and the magnetic field. The maximum increment of oxygen concentration drops to 0.16% when the gas temperature rises to 343 K, and drops to 0.09% when the maximum product of magnetic flux density and gradient is reduced to 101 T^2/m from 563 T^2/m.