Genovoxx Technology (AnyGene.TechnologyTM)
Massively parallel sequencing by synthesis of nucleic acids
Genovoxx is developing a technology for the massively parallel sequencing by synthesis of nucleic acids. This technology enables the sequencing of many millions of nucleic acids molecules in parallel format at the single molecule level.
How ultra fast sequencing by synthesis works
Probe fixation: Randomly distributed fixation and priming of millions of individual DNA molecules.
After fragmentation and addition of an universal primer binding site, genetic material (DNA) is randomly distributed, fixed and primed on the surface of the proprietary AnyDot.ChipTM. Distance between neighbouring molecules averages 1 µm. The sample is applied by simple liquid exchange within a micro fluidic system. Each mm² contains 1 million single DNA molecules ready for sequencing.
Step 1: Polymerase catalyzed incorporation of fluorescently labelled AnyBase.NucleotidesTM.
After removal of unbound DNA fragments, a solution containing polymerase and proprietary AnyBase.NucleotidesTM is applied to the chip. When incorporated into the primer-DNA hybrid, AnyBase.NucleotidesTM cause a reversible stop of the primer-extension (terminating property of nucleotides). Therefore, this step represents a single-base extension. During the stop, incorporated bases on the surface can be detected.
Step 2: Detection of signals on the AnyDot.ChipTM.
Fluorescent dots are detected by the fluorescence detection system. At present, Genovoxx uses a modified state-of-the-art fluorescent microscope. With its proprietary adjustment system, Genovoxx assures that each single fluorescence signal can be properly tracked over the complete sequencing cycles.
Step 3: Removal of terminating property and fluorescent label.
The terminating property and fluorescent label of the incorporated AnyBase.NucleotidesTM are removed completely afterwards. The nucleotides are now extendable similarly to native nucleotides. Thus, the artificial stop of the sequencing process is reversed and the next incorporation cycle can be started.
Cyclic repetition of steps 1 to 3.
For generating sequence data that can be compared with a reference database (for instance NCBI), the length of the sequence snippets has to exceed 15-20 nucleotides. Therefore, steps 1 to 3 are repeated until the majority of all single molecules reaches the required length. This will take, in average, 2 offers of nucleotide incorporations per base.