Isolation of Microsatellite Markers in the Common Carp (Cyprinus carpio)
LOU Yun-Dong Wei Dong-Wang
Abstract
LOU Yun-Dong Wei Dong-Wang
Abstract
A partial common carp genomic library was constructed.45 positive clones were isolated from screening about 2000 clones of the genomic library with a (CA) 15 probe labelled at the 5 end with γ[32]P-ATP.Sequencing of these clones was performed with automated sequencer,and 22 microsatellites were isolated.17 primers were designed based on unique sequences flanking each motif with the software Primer3.PCR on Cyprinus carpio haematopterus was carried out with these primers,and all gave expected bands.Annealing temperature of these primers was between 50 ℃ and 53 ℃.
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A partial common carp genomic library was constructed.45 positive clones were isolated from screening about 2000 clones of the genomic library with a (CA) 15 probe labelled at the 5 end with γ[32]P-ATP.Sequencing of these clones was performed with automated sequencer,and 22 microsatellites were isolated.17 primers were designed based on unique sequences flanking each motif with the software Primer3.PCR on Cyprinus carpio haematopterus was carried out with these primers,and all gave expected bands.Annealing temperature of these primers was between 50 ℃ and 53 ℃.
Key concepts: Cyprinus, Common carp, Carp, Microsatellite, Biology, Genetics, genomic DNA, Genomic library