Soybean cDNA library Gm-c1005
| 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 the apical shoots of 9 to 10 day old etiolated seedlings. The shoot tips including any emerged leaves were harvested for mRNA isolation. The cDNA library was prepared using the Stratagene pBluescript II XR cDNA library construction kit. 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 XL10-Gold host cells. |
| Williams 82 |
| Apical shoot tips, 9-10 day old etiolated seedlings |
| 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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