Soybean cDNA library Gm-c1040
| 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 from hypocotyl and plumule tissues of seeds germinated for three days of the cultivar Williams 82. Complementary DNA was synthesized from mRNA using a poly (dT) primer with a NotI restriction site. EcoR I adapters were ligated to the blunt-ended cDNA fragments followed by digestion with EcoR I and NotI. The cDNA fragments were directionally cloned into the EcoR I- NotI restriction site of the pT7T3-Pac vector. The ligated cDNA fragments were transformed into DH10B host cells (Gibco BRL). |
| Williams 82 |
| Hypocotyl and Plumule, germinating seeds |
| The cDNA library was prepared using the pT7T3-Pac vector (pT7T3, Pharmacia) 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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