454 Life Sciences™ has developed a revolutionary technology, producing millions of raw bases per hour on a single instrument. 454 has also developed system specific software enabling mapping or de novo assembly for whole genome shotgun sequencing of genomes up to 50 megabases. Many biologically meaningful and complex regions of genomes can be analyzed with this system without the time or cost constraints of current DNA sequencing methods. Through this technology 454 Life Sciences provides an enabling solution for ultra-high-throughput DNA sequencing.
Perform Rapid, efficient amplification and sequencing in picoliter format with massive parallelization.
- Fast
Sequence more than 20 million bases per 4.5-hour instrument run. - Cost-Effective
Benefit from reduced cost per base compared to conventional Sanger technology. - Simple
Perform sequencing runs with an easy-to-use instrument requiring minimal steps. - Efficient
Sequence a typical bacterial genome in days with one person - without cloning and colony picking. - Convenient
Use the complete system solution - from sample preparation to data mapping or assembly.
The System:
The instrument is at the heart of the 454 technology. It contains the fluidics and optical systems required to perform and record the sequencing run. In one instrument run sequence a minimum of 20 million base pairs in 4.5 hours.
The PicoTiterPlate™ Device
The PicoTiterPlate Device is a revolutionary advancement that has enabled the 454 technology. The fiber optic plate transmits the chemilluminescent signal, generated by the sequencing reaction, to be recorded by the CCD camera in the instrument.
The Process:
The 454 Sequencing™ process is a start to finish solution. Starting with long or short DNA fragment the 454 process involves library preparation, amplification onto beads and sequencing. The whole process, start to finish, can take one researcher less than a week to complete.
DNA Library Preparation:
Starting with whole genomic DNA to 50 base pair fragments, generate a library that can supply hundreds of sequencing runs of the 454 platform.
Emulsion based PCR (emPCR™):
EmPCR enables the amplification of a DNA fragment immobilized on a bead from a single fragment to 10 million identical copies. This amplification is necessary to generate sufficient identical DNA to obtain a strong signal from the sequencing reaction.
Sequencing:
The sequencing reaction that takes place in the 454 instrument involves the sequential flow of nucleotides and a combination of proprietary enzymes that convert the chemicals generated during a nucleotide incorporation into a chemilluminescent signal that can be detected by CCD camera.
The Software:
The proprietary software that was designed for the 454 platform enables data processing of the chemilluminescent signal produced by the sequencing reaction. The included data analysis software enables whole genome sequencing for de novo or resequencing projects.
