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Mission
The Biotechnology Support Facility falls under the auspices of the Department of Biochemistry and Molecular Biology. The mission of the Biotechnology Support Facility is to “Advance the Science of KUMC Investigators” by
- Providing KUMC investigators with state-of-the-art technologies, methodologies, and instrumentation in a centralized facility that enhances their research.
- Freeing KUMC investigators to intellectually advance their science by performing laborious bench work with automated instrumentation.
- Saving KUMC investigators valuable research time by offering a productivity and performance level that provides superior products and data in less time than they can provide themselves.
- Serving KUMC investigators as an academic resource center for edification in the technology, methodology, and instrumentation of a modern biotechnology core facility.
Available Services
Downloadable order forms for the offered services can be obtained by double clicking on the words “Order form” which are next to the listed services. The order forms are .PDF files which can be filled out online using Adobe Acrobat Reader. If you have problems completing the form online, then you may print the file and fill it out manually. Please contact the facility directly, Tel. (913) 588-6927, E-mail: bwisdom@kumc.edu, for information regarding microarray gene expression analysis or any of our other services.
- DNA Sequencing - Order form
- Oligonucleotide Synthesis - Order form
- Microarray Gene Expression Analysis - Order form
- Peptide Synthesis - Order form
- Amino Acid Analysis - Order form
- Protein/Peptide Sequencing - Order form
Description of Services
DNA Sequencing
DNA sequencing is performed using an ABI 3130XL Genetic Analyzer. The system utilizes dye terminator fluorophores to sequence single or double stranded plasmid templates, BAC templates, or PCR fragments. The contiguous sequence of DNA bases is generated using proprietary base calling software. To enhance sequencing results, the facility recommends Qiagen or similar cartridge purified templates and purified custom primers. Standard sequencing primers are provided by the facility at no additional cost.
Oligonucelotide Synthesis
Oligonucleotide synthesis is performed on an ABI 394-08 DNA/RNA Synthesizer. Synthesis is accomplished on controlled pore glass supports in a 3' to 5' direction. Scale of synthesis on the 394-08 is 40-50 nanomole, 200 nanomole, or 1 micromole. The BSF recommends purified oligonucleotides where the purity of the primer may enhance the investigator's results. Purification of oligonucleotides is performed on single-use trityl specific cartridges.
Microarray Gene Expression Analysis
The BSF oversees and operates the School of Medicine Microarray Core Facility, which provides gene expression analysis using the Affymetrix GeneChip Instrumentation System or Affymetrix 428 Array Scanner for custom glass slide microarrays. The Affymetrix GeneChip Instrumentation System includes an Affymetrix GeneChip Scanner 3000, GeneChip Fluidics Station 450, and GeneChip Hybridization Oven 640. An Agilent 2100 Bioanalyzer provides rapid quality assessment of total RNA submissions and target preparations used in gene expression analyses.
Peptide Synthesis
Peptide synthesis is performed on a Rainin Symphony Quartet Peptide Synthesizer. This instrument is capable of simultaneously synthesizing four peptides using Fmoc chemistry. Routine levels of synthesis are 25 and 50 micromole scales. Multiple Antigenic Peptides (MAPs) can also be synthesized with the Quartet by using a 4 or 8-branched immunologically inert lysine core resin.
Amino Acid Analysis
Amino acid analysis is performed on a Hitachi L-8800 Amino Acid Analyzer. Protein or peptide hydrolysates are separated into their constituent amino acids by reverse phase HPLC and post column derivatized with ninhydrin. Derivatized amino acids are quantitated and their ratio relative to one another is determined based on a known amino acid standard. Samples must be submitted in a volatile liquid or lyophilized form. Blotted (PVDF) samples are not recommended as the membrane may interfere with hydrolysis. Removal of all salts, buffers, and/or detergents prior to analysis will enhance amino acid recoveries and accuracy of data.
Protein/Peptide Sequencing
Protein or peptide sequencing is performed on the facility's ABI Procise 491 Protein Sequencer using standard Edman degradation chemistry. This chemistry derivatizes and removes one amino acid at a time from the amino-terminal end of the protein or peptide that is being sequenced. The amino acid is then subjected to on-line HPLC. The liberated amino acid is identified by its retention time when compared to retention times of standard amino acids. Samples for sequencing must be submitted in a liquid, lyophilized, or blotted membrane (PVDF) form. Samples submitted for sequencing must be free of salts, buffers, or detergents that will impair the sequencing results.
Staff
Director: Dr. Gerald M. Carlson, Chairperson Biochemistry and Molecular Biology
Lab Manager: Bo Wisdom, M.A.
Research Associates: Jim Rouse and Clark Bloomer
Research Assistant: Rosanne Skinner
For Further Information Contact
Bo Wisdom
Biotechnology Support Facility
5022 WHW
University of Kansas Medical Center, (KUMC)
3901 Rainbow Blvd.
Kansas City, KS 66160
Phone: (913) 588-6927
FAX: (913) 588-7131
E-mail: bwisdom@kumc.edu or cbloomer@kumc.edu
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