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Sample Preparation
INT's approach to identification requires a unique approach to sample preparation. The resulting method is robust and rapid at providing clean concentrated nucleic acid or protein from a wide range of sample types. The process works as follows:
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The sample preparation system works with a wide variety of sample types, including liquids, solids, spores, tissue, blood, and insects. It can also process dirty samples, such as soil and sand. The user introduces a sample into the cartridge then snaps the loaded cartridge into a fitted seat on the sample prep drive device. |
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The user presses the start button to begin operation and the remaining process is automated. The cartridge includes an internal rotating section in which the sample sits. This rotating chamber moves the sample through each step. |
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Next, the device must release the DNA or RNA from the cells in the sample. The sample is moved to a chamber in the cartridge that is filled with a lysis buffer and 100 micron glass beads (the larger objects in the image at left). |
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An ultrasonication device within the analyzer vibrates the glass beads which then break open the sample cells releasing the nucleic acid. The ultrasonication also breaks the DNA and RNA down to usable lengths. |
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Nano-paramagnetic clusters in the lysis buffer bind to the nucleic acids. These particles are only 10-25 nanometers in diameter. |
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An electromagnet in the drive device concentrates the para-magnetic particle-DNA cluster to the bottom of the sample chamber. A built-in 'syringe' in the disposable pulls the concentrated nucleic acid from the chamber through a filter to the denaturation chamber. The electromagnets hold the nucleic acids in a channel while a wash buffer is flowed over the nucleic acids to wash away contaminants before analysis. |
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The cleaned and concentrated nucleic acid is released from the particles into an amplification buffer and denatured by heating. If needed, the device can heat and cool the sample rapidly to amplify the number of nucleic acid sequences by PCR to increase the working material available. |
Our sample preparation technology has been used to prepare DNA, RNA and protein from a variety of samples including bacteria, viruses, plant material, insects, and animal and human samples. The process removes inhibitors, thus providing clean material from complex sample matrices including blood, tissue, soil and air filter washes. The process shows significant advantages over competing approaches, with a recovery of better than 90% of nucleic acids over a range of femtograms to hundreds of micrograms of input material per sample. In addition, the process allows for the production of nucleic acids with tight size distributions for use in automated gene sequencers. INT is preparing sample preparation kits to bring to market and developing automated systems for field use and high throughput sample preparation.