Soybean cDNA library Gm-c1063
| Dr. Randy C. Shoemaker | Phone: | 515 294 6233 | e-mail: | rcsshoe@iastate.edu | ||||
| Department of Agronomy | FAX: | 515 294 2299 | ||||||
| USDA-ARS | ||||||||
| G401 Agronomy Hall | ||||||||
| Iowa State University | ||||||||
| Ames, IA 50011-1010 |
| The cDNA library was constructed from mRNA isolated germinating shoots of the cultivar Williams. The seeds were allowed to germinate for 24 hours prior to harvesting the germinating shoots. Complementary DNA was synthesized from mRNA using a primer consisting of a poly (dT) sequence with a Xho I restriction site. EcoR I adapters were ligated to the blunt-ended cDNA fragments followed by Xho I digestion. The cDNA fragments were directionally cloned into the EcoR I-Xho I restriction site of the pBluescript vector. The ligated cDNA fragments were transformed into DH10B host cells (Gibco BRL). |
| Williams |
| Germinating shoot, 24 hour germination |
| The cDNA library was prepared using the pBluescript II SK (+) vector (Stratagene) to generate a plasmid library. The vector provides ampicillin resistance and allows blue-white color selection for recombinant phagemids. The cloned cDNA fragment can be amplified by the polymerase chain reaction using one of the following vector primer pair combinations: T3 and T7 or M13 universal and M13 reverse. |
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