Integrated on-line system for DNA sequencing by capillary electrophoresis: From template to called bases
- Ames Lab., IA (United States)
An integrated on-line prototype for coupling a microreactor to capillary electrophoresis for DNA sequencing has been demonstrated. A dye-labeled terminator cycle-sequencing reaction is performed in a fused-silica capillary. Subsequently, the sequencing ladder is directly injected into a size-exclusion chromatographic column operated at nearly 95{degree}C for purification. On-line injection to a capillary for electrophoresis is accomplished at a junction set at nearly 70{degree}C. High temperature at the purification column and injection junction prevents the renaturation of DNA fragments during on-line transfer without affecting the separation. The high solubility of DNA in and the relatively low ionic strength of 1 x TE buffer permit both effective purification and electrokinetic injection of the DNA sample. The system is compatible with highly efficient separations by a replaceable poly(ethylene oxide) polymer solution in uncoated capillary tubes. Future automation and adaptation to a multiple-capillary array system should allow high-speed, high-throughput DNA sequencing from templates to called bases in one step. 32 refs., 5 figs.
- DOE Contract Number:
- W-7405-ENG-82
- OSTI ID:
- 466326
- Journal Information:
- Analytical Chemistry (Washington), Vol. 69, Issue 4; Other Information: PBD: 15 Feb 1997
- Country of Publication:
- United States
- Language:
- English
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