Soybean cDNA library Gm-c1042
| 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 2 week old seedlings with the cotyledons removed at the time of harvest. The seedlings for the cultivar Raiden were grown in a growth chamber using germination paper. 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). |
| Raiden |
| Whole seedling without cotyledons |
| 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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