Abstract
This article reports the development of microsatellite primers for giant freshwater prawns (Macrobrachium rosenbergii), using the genome library for this type of prawns and biotinylated probe in isolating six types of DNA fragments with base sequence repeats, namely (AG)10, (TG)10, (CAA)10, (CAG)10, (GAT)10 and (TAC)10. Four clones harboring microsatellites were chosen, namely SH2-9F, SH2-10C, SH2-11D and SH3-11G. Base sequencing found no microsatellite sequences. Four pairs of primers were designed, namely DTLSH 7, DTLSH 8, DTLSH 9 and DTLSH 12, respectively. These primers were tested on the DNA of giant freshwater prawns, and DNA band sizes were found to be 131, 174, 210 and 193 per 205 bp, respectively. The developed microsatellite primers may be used in conjunction with primers for other types of prawns. Genetic diversity checking of giant freshwater prawns in Thailand showed that their population in Thailand may be classified into two groups: one with the closest genetic relationship being those from Khon Kaen, Samut Songkhram and Ang Thong provinces while the other with similar genetic characteristics were those from these three provinces as compared to those from Surat Thani province.
Highlights
Macrobrachium rosenbergii, or its generic name the giant freshwater prawn, is the freshwater prawn popularly raised in India, in the Southeast Asian countries, in Northern Australia and in certain parts of the Pacific and Indian Oceans
Comparing with lambda DNA, which was used as a marker, the DNA concentration (SH) was approximately 50 ng/μl, which were sufficient for test in the step
After the genomic DNA of the prawns was digested with the restriction enzyme Tru 9I or MseI and ligated with MseI-Adaptor, it was found that the prawn DNA samples were on all three A-lanes
Summary
Macrobrachium rosenbergii, or its generic name the giant freshwater prawn, is the freshwater prawn popularly raised in India, in the Southeast Asian countries, in Northern Australia and in certain parts of the Pacific and Indian Oceans. In Thailand, the aquaculture part is considered an industry with great significance to the country as prawns can be a principal aquacultural protein source that is high in nutritional values. They are one of Thailand’s top export goods in terms of income earnings. Development of microsatellite markers will, be very useful in identifying polymorphisms and even in the mapping of a sample population from the same species. Thailand’s giant freshwater prawns have been classified in this latter group, which has so far had 11 pairs of microsatellite primers [4]. Its data base for development of additional microsatellite primers to cover other parts of the prawns’ genome would maximize the usage and accuracy in the breeding programs in the future
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