Array based
sequencing-by-synthesis
As well as its proprietary real-time single molecule sequencing system, Mobious has pursued its own, novel approach to sequencing by synthesis. We have developed applications that will upgrade conventional biochip arrays into highly paralleled sequencing systems.
Single molecule sequencing is our goal, as the potential throughput and cost would make it the platform of choice for high throughput, genome wide sequencing. In its current form however, the cost and complexity of single molecule sequencing approaches puts it out of reach for routine genomic analysis. Mobious is in a position to bridge this gap via a novel and powerful approach to sequencing-by-synthesis.
The general approach to polynucleotide sequencing is known as sequencing-by-synthesis (SBS). Two main approaches have been exploited for SBS so far. These are Labelled Nucleotide approach and others using the detection of pyrophosphate.
Both of these techniques have drawbacks. The use of labelled nucleotides can interfere with the ability of the polymerase molecule use to “grow” the new sequencing strand. This results in significant issues relating to read length and data integrity.
SBS systems that rely on the detection of pyrophosphate as a marker for successful nucleotide addition, suffer drawbacks related to the complexity of this “signal chain”. Apart from the polymerase enzyme, which is essential for the sequencing process, pyrophosphate detection systems need at least another two enzymes, Sulfurylase and Luciferase. These enzymes have different environmental requirements and have to be located at a distance from the site of pyrophosphate generation. Such factors affect the data integrity of the system.
Mobious’s proprietary approach to SBS circumvents the use of labelled nucleotides and secondary enzyme transduction systems. Known as Polykinetic Sequencing™, it takes advantage of the natural selective mechanisms present with the polymerase reaction mechanism.
For a polymerase to add a nucleotide to a growing DNA chain, it must first bind to the strand and then “select” a nucleotide from solution in order to “test” whether or not the nucleotide is a correct base or a “match”.
If a “match” is confirmed, then the polymerase enzyme proceeds to add the base to the growing DNA molecule. If the base, however, turns out not to “match” then it is returned to solution.
These two sequences, match and mismatch, involve different periods. A match results in the base being added to the growing strand, which this involves several “time consuming” steps and it is this time difference which is exploited in Mobious’s sequencing-by-synthesis approach. It places the polymerase in a flow, adds a single nucleotide at a time and derives sequence data by measuring the amount of time the polymerase attaches to the growing strand. A number of detection strategies may be applied to the technique. By simply labelling the polymerase with a fluorescent species, the approach can be applied in the context of conventional array formats.
In conjunction with Mobious’s proprietary microfluidic technologies, therefore, the polykinetic sequencing-by-synthesis can turn a DNA array into a high throughput genomic sequencer.
As well as its proprietary real-time single molecule sequencing system, Mobious has pursued its own, novel approach to sequencing by synthesis. We have developed applications that will upgrade conventional biochip arrays into highly paralleled sequencing systems.
Single molecule sequencing is our goal, as the potential throughput and cost would make it the platform of choice for high throughput, genome wide sequencing. In its current form however, the cost and complexity of single molecule sequencing approaches puts it out of reach for routine genomic analysis. Mobious is in a position to bridge this gap via a novel and powerful approach to sequencing-by-synthesis.
The general approach to polynucleotide sequencing is known as sequencing-by-synthesis (SBS). Two main approaches have been exploited for SBS so far. These are Labelled Nucleotide approach and others using the detection of pyrophosphate.
Both of these techniques have drawbacks. The use of labelled nucleotides can interfere with the ability of the polymerase molecule use to “grow” the new sequencing strand. This results in significant issues relating to read length and data integrity.
SBS systems that rely on the detection of pyrophosphate as a marker for successful nucleotide addition, suffer drawbacks related to the complexity of this “signal chain”. Apart from the polymerase enzyme, which is essential for the sequencing process, pyrophosphate detection systems need at least another two enzymes, Sulfurylase and Luciferase. These enzymes have different environmental requirements and have to be located at a distance from the site of pyrophosphate generation. Such factors affect the data integrity of the system.
Mobious’s proprietary approach to SBS circumvents the use of labelled nucleotides and secondary enzyme transduction systems. Known as Polykinetic Sequencing™, it takes advantage of the natural selective mechanisms present with the polymerase reaction mechanism.
For a polymerase to add a nucleotide to a growing DNA chain, it must first bind to the strand and then “select” a nucleotide from solution in order to “test” whether or not the nucleotide is a correct base or a “match”.
If a “match” is confirmed, then the polymerase enzyme proceeds to add the base to the growing DNA molecule. If the base, however, turns out not to “match” then it is returned to solution.
These two sequences, match and mismatch, involve different periods. A match results in the base being added to the growing strand, which this involves several “time consuming” steps and it is this time difference which is exploited in Mobious’s sequencing-by-synthesis approach. It places the polymerase in a flow, adds a single nucleotide at a time and derives sequence data by measuring the amount of time the polymerase attaches to the growing strand. A number of detection strategies may be applied to the technique. By simply labelling the polymerase with a fluorescent species, the approach can be applied in the context of conventional array formats.
In conjunction with Mobious’s proprietary microfluidic technologies, therefore, the polykinetic sequencing-by-synthesis can turn a DNA array into a high throughput genomic sequencer.