PERTANIKA JOURNAL OF TROPICAL AGRICULTURAL SCIENCE

 

e-ISSN 2231-8542
ISSN 1511-3701

Home / Regular Issue / JTAS Vol. 28 (1) Apr. 2005 / JTAS-0304-2005

 

Rapid and Non-radioactive Detection Method of Microsatellites in Mystus nemurus: A Refined Technique

Chan Soon Choy, Siti Shapor Siraj, Tan Soon Guan and Khatijah Yusoff.

Pertanika Journal of Tropical Agricultural Science, Volume 28, Issue 1, April 2005

Keywords: Microsatellites, Mystus nemurus, cloning, polymerase chain reaction (PCR), random amplified microsatellites (RAMs)

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A simple and rapid method of DNA microsatellite isolation based on the Random Amplified Microsatellites (RAMs) PCR technique was used in this study. The work presented here is part of a continuous effort in refining and perfecting the technique for more rapid, effective and optimum productivity in single locus microsatellite marker development for the River catfish, Mystus nemurus. The current refined protocol for microsatellite isolation was able to detect a total of 135 microsatellite regions resulting in 42 unique genomic sequences being submitted to GenBank. This refined technique is able to reduce the total time required from PCR cloning till sequencing specific microsatellite regions to less than three and a half months