| DNA Sequencing |
| DNA Synthesis |
| Biotechnology Consult |
| Biopolymer Links |
| Staff |
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| DNA SEQUENCING: |
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Instrumentation |
The Biopolymer Laboratory performs DNA sequencing analysis using the automated sequencer model ABI 373XL from Perkin Elmer/Applied Biosystems. DNA sequencing reactions are carried out using the ABI Big Dye Terminator cycle sequencing reagent. |
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Materials required |
DNA templates (single stranded vector, double stranded vector and PCR fragment) are provided by the user. All templates must be carefully purified in order to remove possible contaminants, because poor template quality is the major source of fluorescent sequencing failure. Typically Qiagen columns give the most reliable DNA for sequencing when compared to other procedures. Three to five microliters of template at a concentration of 0.2 microgram/microliter in pure water are needed for analysis. The use of Tris-EDTA buffer should be avoided as this can interfere with the sequencing reaction. Prior to sequencing, a brief analysis of the DNA templates on mini agarose gel is strongly recommended to confirm the adequacy of the preparation. Standard M13 primers are provided at no charge by the Biopolymer Laboratory. Custom primer can be ordered by using an oligonucleotide request form. All primers are used at the concentration of 1 picomole/microliter in pure water. Five to ten microliters of primer are needed for each sample. |
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| Results |
Color printouts of analyzed chromatograms are provided for each successful reaction. Sequencing results are stored on diskettes. Typically, sequences of 500 to 700 accurate bases can be obtained from clean DNA templates. |
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| DNA SYNTHESIS |
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Instrumentation |
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The Biopolymer Laboratory has two DNA synthesizers: an Applied Biosystems Model 381A and a Beckman Model 1000M. The ABI 381A is a single column dedicated to the synthesis of speciality DNA. The Beckman 1000M (Oligo 1000M DNA Synthesizer) is a fully automated synthesizer with a built-in Trityl monitor and extensive diagnostic functions to monitor the synthesis process and ensure the quality and purity of the synthesized oligonucleotides. It has the capability to synthesize eight oligonucleotides sequentially in a rapid and economical cycle time.
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| BIOTECHNOLOGY CONSULT |
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University of South Alabama investigators are invited to consult with Dr. Tin Cao, Director of the Biopolymer Laboratory, regarding experimental design, biotechnical problems and alternative strategies related to their molcular biology research. In addition, the Biopolymer Laboratory is prepared to perform specialized procedures on an as needed basis. |
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| BIOPOLYMER LINKS |
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| BIOPOLYMER STAFF |
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Dr. Tin Cao, Director
Andrea Britain, Research Technologist II
Telephone: (251) 460-7264
FAX: (251) 460-7931
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