Soybean cDNA library Gm-c1084
| Dr. Randy C. Shoemaker (Madan Bhattacharyya) | 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 by M. Bhattacharyya from mRNA isolated from etiolated hypocotyls from the cultivar Williams 82. Tissue was inoculated with Phytophthora sojae race 1 and tissues were harvested 2 and 4 hours following infection. The library is the pool of these two time points. 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 82 |
| Etiolated hypocotyls (Williams 82) inoculated with Phytophthora sojae race 1 and tissues harvested 2 and 4 hours following infection. Library is the pool of these two time points. |
| 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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