[Enrichment of giant panda microsatellite markers using dynal magnet beads].
The 400 -600 bp DNA fractions of giant panda containing STR sequences were captured by hybridization with the oligonucleotide probes attached to streptavadin coated magnetic beads (Dynal). The enriched DNA were ligated into pGEM-T and then transformed into E. coil JM109 competent cells. In total 260 positive clones were identified from 2 880 transformants in the libraries which were screened by gamma-32 P radiolabelled probes. Finally, we got 54 sequences and successfully designed 37 pairs of STR primers for giant panda. The results showed that this method is very efficient to isolate microsatellite markers.
Shen
Fu-Jun
FJ
Chengdu Research Base of Giant Panda Breeding, Key Lab for the Conservation Genetics & Reproduction of Sichuan Province, Chengdu 610081, China. shenfuj@263.sina.com
Watts
Phill
P
Zhang
Zhi-He
ZH
Zhang
An-Ju
AJ
Sanderson
Stephanie
S
Kemp
Steve J
SJ
Yue
Bi-Song
BS
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers; D015723:Gene Library; D008280:Magnetics; D018895:Microsatellite Repeats; D008863:Microspheres; D009693:Nucleic Acid Hybridization / Q000379:methods; D011110:Polymorphism, Genetic; D020080:Tandem Repeat Sequences; D001503:Ursidae / Q000235:genetics*
D017931:DNA Primers