Soybean cDNA library Gm-c1087


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. Joe Polacco Phone:   573-882-4789       e-mail:   PolaccoJ@missouri.edu
Biochemistry Department FAX: 573-882-5635    
112 Schweitzer Hall        
University of Missouri-Columbia        
Columbia, MO 65211        

Short Description of Library:
The mRNA was prepared by Joe Polacco, University of Missouri, using the PolyAtract mRNA isolation system from PROMEGA. The cDNA was prepared using the STRATAGENE kit. Complementary DNA was synthesized from mRNA using a primer consisting of a poly (dT) sequence with an XhoI restriction site. Eco RI adapters were ligated to the blunt-ended cDNA fragments, followed by Xho I digestion. cDNA was size fractionated using a drip column containing Sepharose CL 2B to release residual linker-primer Eco RI adapters and small cDNA fragments. The cDNA fragments were directionally cloned into the Eco RI-Xho I restriction site of the pBlueScript II SK+ vector (previously dephosphorylated). The ligated cDNA fragments were transformed into DH10B by electroporation. Million-fold amplified stock is stored at -80° in 7% DMSO.

Genotype/Cultivar:
Williams 82

Organ/Tissue:
Root 11 days old after germination in hydroponic system, without phosphate (time of maximal acid phosphatase activity). Roots were directly immersed in aerated nutrient solution.

Vector Information:
pBluescript" II SK(+)
# independent transformants: 9 x 105 cpu
# proportion of white colonies: 80%
# colonies checked for insert: 37 (26 white + 11 blue)
24/26 white with insert (average size 800 bp) (Range = 400-1,700 bp)
5/11 blue with insert (1 = 400 bp; 4= 100 bp)
Colonies with real insert, i.e. > 100 bp, = 65%
# independent transformants with insert: 5.8 x 105 cpu
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