Development of EST-SSRs by an efficient FIASCO-based strategy: a case study in rare minnow (Gobiocyrpis rarus).
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.
Cheng
L
L
The State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, the Chinese Academy of Sciences, Wuhan, China.
Liao
X
X
Yu
X
X
Tong
J
J
D000483:Alleles; D054458:Amplified Fragment Length Polymorphism Analysis; D000818:Animals; D001483:Base Sequence; D016000:Cluster Analysis; D003530:Cyprinidae / Q000235:genetics*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D020224:Expressed Sequence Tags; D005819:Genetic Markers; D014644:Genetic Variation; D018895:Microsatellite Repeats / Q000235:genetics; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic
D018076:DNA, Complementary; D005819:Genetic Markers