Soybean cDNA library Gm-c1037
| Dr. Lila O. Vodkin | Phone: | 217 244 6147 | e-mail: | l-vodkin@uiuc.edu | ||||
| University of Illinois at Champaign-Urbana | ||||||||
| Department of Crop Science | ||||||||
| 384 Edward R. Madigan Laboratory | ||||||||
| 1201 W. Gregory Drive | ||||||||
| Urbana, IL 61801 |
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The cDNA library was constructed from mRNA isolated from fully expanded leaves of greenhouse grown plants that were 2 week old.The library was prepared using the Life Technologies pSuperScript cDNA library construction kit. Complementary DNA was synthesized from mRNA using a poly (dT) sequence with a Not I restrictions site. Sal I linkers adapters were ligated to the blunt-ended cDNA fragments followed by Not I digestion. The cDNA fragments were directionally cloned into the Not I-Sal I restriction site of the pSPORT 1 vector. The ligated cDNA fragments were transformed into E. coli ElectroMax DH10B host cells. This record shows additional ligation. Transformation efficiency: 105 Percent white colonies: 98% Average insert size of white colonies: 850 bp |
| Williams |
| Fully expanded leaves of 2 week old greenhouse grown plants |
| The cDNA library was prepared using the pSPORT 1 vector (Life Technologies) to generate a plasmid library. The vector provides ampicillin resistance and allows blue-white color selection for recombinant plasmids. The vector also contains unique Sal I and Not I sites needed to clone cDNA directionally. The cloned cDNA fragment can be amplified by the polymerase chain reaction using one of the following vector primer pair combinations: M13 universal and M13 reverse or T7 and SP6. |
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