The application of ultrasound as a rapid method to provide DNA fragments suitable for detection by DNA biosensors.
Contamination of food and water supplies by microorganisms such as Escherichia coli, the need for point-of-care bedside analysis of biological samples, and concerns about terrorist attacks using biological organisms, have made the development of fast, reliable, and sensitive analytical methodologies for use in monitoring of pathogens very important. With a variety of biosensors being developed for extremely sensitive and rapid nucleic acid diagnostics, it has become even more important to shift focus towards creation of methods to decrease the amount of time and effort necessary for sample preparation. The application of ultrasound has the potential to create DNA fragments from genomic material with lengths that are suitable for determination using biosensors and microarrays. For example, application of 85 W power at a frequency of 20 kHz can produce a preponderance of fragments of 100-400 base pairs (bp) within several seconds, and sample processing can lead to over 75% conversion from genomic material to fragments in times of 20-30 s. A proportion of these fragments are in a single-stranded state and are suitable for hydridization with immobilized single-stranded DNA probe oligonucleotides using a fiber optic biosensor. Control of factors such as salt concentration, exposure time, ultrasound power, and the initial temperature of the solution, can affect the length and form (single- or double-stranded) of DNA fragments that are generated by ultrasound, and average fragment length can be adjusted by selection of these operating parameters.
Mann
Tina L
TL
Chemical Sensors Group, Department of Chemistry, University of Toronto, 3359 Mississauga Road North, Mississauga, ON, Canada L5L 1C6.
Krull
Ulrich J
UJ
D015374:Biosensing Techniques / Q000379:methods*; D004247:DNA / Q000032:analysis / Q000737:chemistry* / Q000528:radiation effects* / Q000648:ultrastructure; D015342:DNA Probes / Q000032:analysis / Q000138:chemical synthesis* / Q000648:ultrastructure; D004307:Dose-Response Relationship, Radiation; D015394:Molecular Structure; D008970:Molecular Weight; D009841:Oligonucleotides / Q000138:chemical synthesis*; D011829:Radiation Dosage; D013010:Sonication
D015342:DNA Probes; D009841:Oligonucleotides; D004247:DNA