Soybean cDNA library Gm-c1073


Clones from this library are available from
    Biogenetic Services
    801 32nd Ave.
    Brookings, SD 57006 USA
    phone: 800 423 4163
    email: info@biogeneticservices.com

To insure that credit is given to the laboratory and scientists donating this valuable resource to the scientific community, the information provided with the library must be maintained and included with any transfer of biological materials or data derived from the library.

Source of Library (Donor):
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        

Short Description of Library:
The cDNA library was constructed from mRNA isolated from 2-3 week old seedlings that were induced for symptoms of SDS (Sudden Death Syndrome) disease by the translocution of culture filtrate of Fusarium solani f. sp. glycines (Plant Cell Report 18:375-380). Cultivar Williams 82 is susceptible to the disease SDS. Plant tissue (expanded leaves, folded leaves, and new shoots) were collected at 1, 6, and 24 hrs after inoculation and their mRNA pooled equally for cDNA construction. The library was prepared using the Stratagene pBluescript II SK (+) library construction kit. Complementary DNA was synthesized from mRNA using a primer consisting of a poly (dT) sequence with an Xho I restriction site. Eco RI adaptors were ligated to the blunt-ended cDNA fragments followed by Xho I digestion. The cDNA insert is protected from Xho I digestion via methylation during first strand cDNA synthesis. The cDNA fragments were directionally cloned into the Eco RI-Xho I restriction site of the pBluescript vector. The ligated cDNA fragments were transformed into E. coli ElectroMax DH10B host cells.

Titer of cDNA clones: 3 x 105 CFU/ml
Percent white colonies: 60%
Average insert size of white colonies: 650 bp

Genotype/Cultivar:
Williams 82

Organ/Tissue:
2-3 week old seedlings, induced with SDS disease

Vector Information:
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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