Public EST Project for Soybean
Record of Deposit
Soybean cDNA Library Gm-c1034
Very young seed coat
The soybean cDNA library is being made publicly available by the following laboratory. 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.
Library Source/Contributor
Dr. Lila O. Vodkin
University of Illinois at Urbana-Champaign
Department of Crop Sciences
384 Edward R. Madigan Laboratory
1201 W. Gregory Drive
Urbana, IL 61801
Phone: 217-244-6147
Fax: 217-333-4777
e-mail: l-vodkin@uiuc.edu
Genotype/Cultivar
Williams
Vector Information
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.
Library Description
The cDNA library was constructed from mRNA isolated from very young seed coats (2050 mgs fresh weight) of greenhouse grown plants. 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.
Transformation efficiency: 10 5colonies/ug vector DNA
Percent white colonies: 95
Average insert size: 830bp
Disposition: These clones are being sent so the plates can be replicated. Send one plate back to
the originating laboratory and one plate to John Martin or Marco Marra at Washington University
for sequencing. All clone identifiers and shipping instructions are to be determined by Genome
Systems and Washington University.
NAME ASSIGNED BY GENOME SYSTEM: Gm-c1034
Information for Genome Systems:
The libraries were mailed on 11/18/99 in two tubes that were labelled Vodkin, Seed coat cDNA
(20-50).
Dilution information: 500 uls of the enclosed dilution (per tube) of the library will give 2000
colonies. Please plate on X-gal/IPTG plates and pick ONLY the white colonies. Initial tests
show <10% blue colonies.
Picking Instructions: These cells are being sent so that colonies can be picked robotically and
entered into 384 well plates for preliminary analysis and eventually for high density filter arrays.
Please pick up to 2000 colonies. Send one set of plates back to my laboratory and keep one set
at Genome Systems.
Disposition: These clones are being sent so the plates can be replicated. Send one plate back to
the originating laboratory and one plate to John Martin or Marco Marra at Washington University
for sequencing. All clone identifiers and shipping instructions are to be determined by Genome
Systems and Washington University.