Development of EST-SSRs by an Efficient FIASCO-Based Strategy: A Case Study in Rare Minnow (Gobiocyrpis Rarus)

Abstract

Type I markers are useful for comparative mapping and other genetic analyses, but relatively difficult to develop. In the present study a microsatellite (SSR)-enriched cDNA library was constructed for the first time using the fast isolation by AFLP of sequences containing repeats (FIASCO) method in a small fish, Chinese rare minnow (Gobiocypris rarus). A total of 97.4% of the expressed sequence tags (ESTs) contained targeted CA-repeats, in which 29 unique EST-SSRs were identified. Ten out of the 28 loci for which primer pairs were designed were polymorphic with alleles ranging from three to seven (mean 4.50). Some of these EST-SSRs can be amplified in other species. These results proved that cDNA-FIASCO is an efficient way to isolate novel EST-SSRs in a fish.

Keywords:

• cDNA library
• Expressed sequence tag
• FIASCO
• Fish
• Microsatellite

The authors thank Dr. J. Wang and Ms. C. You for collection of fish samples. This study was supported by the Department of Science and Technology of China (2004CB117405), Natural Science Foundation of China (30370225), and Department of Science and Technology of Hubei Province (2004ABC005).