This project's objective was to develop an automated method for DNA extraction, purification, concentration, and quantification from a complex sample.
The tabletop device developed is fully automated, uses two different detectors for the DNA plug and the focusing interface, delivers the DNA extract into a fresh tube for downstream analysis, and extracts DNA from both clean and dirty buccal swabs in approximately 5 minutes. The device was tested with a dirty buccal swab or a buccal swab completely covered in soil. The device can be used in field lab settings and can be operated by minimally trained personnel with a software package. Further testing of the device indicates that the extraction efficiency can be improved by applying a negatively charged coating to the capillary, performing longer extractions, and extracting DNA from the same sample multiple times. In addition, the device can extract DNA from sample volumes as low as 5 ml. GEITP (gradient elution isotachophoresis) data indicate that DNA has been extracted from samples such as bone, gum, and plant material matrices. GEITP is a combination of gradient elution moving boundary electrophoresis and isotachophoresis. 89 figures, 12 tables, and 10 references