Identification of abundant and informative microsatellites from shrimp (Penaeus monodon) genome.
Microsatellites were isolated from P. monodon genomic libraries by direct sequencing of recombinant clones without probe screening. Forty-nine out of 83 clones sequenced contained 99 microsatellite arrays of three or more repeats. When five or more and ten or more repeats were considered, 28 and 14 microsatellites were detected, respectively. The 99 microsatellites were classified as perfect (75%), imperfect (6%), compound perfect (3%) and compound imperfect (16%). The abundance of di-, tri-, tetra- and hexanucleotide repeats were 67%, 20%, 9% and 3%, respectively. The dinucleotide repeats included 36 (CT)n, 31 (GT)n, 17(AT)n and 3 (CG)n. One octanucleotide repeat (ATTTATTC)5 was found within a large repeat sequence. Optimal annealing temperatures were determined for PCR using 11 primer sets encompassing 15 microsatellites. Ten primer sets provided successful amplifications with allele sizes generally ranging from 139 to 410 bp. All these primers amplified polymorphic loci with PIC values ranging from 0.63 to 0.96. Two primer sets amplified additional bands which can easily be distinguished from the bands of the main locus. Three out of 10 P. monodon microsatellites also amplified alleles in P. vannamei. The abundance and informative nature of P. monodon microsatellites and their potential for cross-species amplification make them useful for genetic studies.
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Department of Environmental and Population Health, Tufts University School of Veterinary Medicine, North Grafton, MA 01536, USA.
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Wyrzykowski
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Alcivar-Warren
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D000818:Animals; D017756:Aquaculture; D004247:DNA / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D015723:Gene Library; D018895:Microsatellite Repeats / Q000235:genetics*; D033561:Penaeidae / Q000737:chemistry / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D011110:Polymorphism, Genetic / Q000235:genetics*; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004247:DNA