Effective isolation of nucleic acids from biological samples is an essential prerequisite for efficient downstream amplification, detection and quantification of specific genetic sequences via quantitative polymerase chain reaction (qPCR). The extraction process requires lysing cells with harsh extraction reagents such as detergents or enzymes, which results in a mixture of nucleic acids, cellular debris and extraction reagents. The nucleic acids are separated from the mixture using a technique such as organic solvent extraction, chromatography, centrifugation or dialysis. These techniques may be time-consuming, and often require multiple washing steps. Up to 15 percent of all molecular biology research time may be spent purifying samples. A simplified method of isolating nucleic acids from a biological sample is needed to reduce the time spent on purification.

UW–Madison researchers have developed a device and method for extracting and purifying a desired fraction from cultured cells, tissue samples and other biological materials. A biological sample, including both non-desired material and a fraction-bound solid phase substrate, is added to an input zone. The input zone is adjacent to a separation zone that includes an isolation buffer. A force moves the fraction-bound solid phase substrate from the input zone, through the separation zone and into an output zone, leaving the non-desired material behind. The improved purification method is simple, more efficient and produces a higher throughput than prior devices and methods. The device may be configured to allow for quantification of the fraction in the biological sample via labeling of the fraction-bound solid phase substrate.

File Number: P110080US01