Information courtesy of Roche 454 Life Sciences
The Genome Sequencer System provides a unique technology for sequencing DNA samples in an efficient, massively parallel and cost–effective manner. The Genome Sequencer FLX System has a number of applications, including de novo sequencing, resequencing of whole genomes and target DNA regions, metagenomics and RNA analysis.
454 Sequencing is based on sequencing–by–synthesis where nucleotides are flowed sequentially in a fixed order across the Pico Titer Plate device during a sequencing run. During the nucleotide flow, hundreds of thousands of beads each carrying millions of copies of a unique single–stranded DNA molecule are sequenced in parallel. If a nucleotide complementary to the template strand is flowed into a well, the polymerase extends the existing DNA strand by adding nucleotides. Addition of one or more nucleotides results in a reaction that generates a light signal that is recorded by the CCD camera in the instrument. The signal strength is proportional to the number of nucleotides incorporated in a single nucleotide flow.
The Pico Titer Plate (PTP)can be divided into a number of different regions allowing the sequencing of a number of different samples on a single PTP. Multiplex Identifiers (MIDs) expand multiplexing options beyond physical separation of samples by introducing a unique 10–bp tag, enabling pooling for simultaneous amplification and sequencing. The sizes of the different regions will dictate the number of reads per region that can be expected.
|Sequencing Kit||Number of Regions||Reads per Region||Run Throughput per Region||Run Throughput per PicoTiterPlate|
|FLX+||2||450,000–650,000||250-400 Mb||500-800 Mb|
|FLX||2||450,000–650,000||180–280 Mb||360–560 Mb|
|FLX+||4||160,000–250,000||160-250 Mb||400-640 Mb|
|FLX||4||160,000–250,000||60-110 Mb||240-440 Mb|
|FLX+||8||80,000–120,000||80-120 Mb||320-560 Mb|
|FLX||8||80,000–120,000||30-55 Mb||240-440 Mb|
|FLX+||16||25,000–40,000||25-40 Mb||240-400 Mb|
|FLX||16||25,000–40,000||10-20 Mb||160-320 Mb|
|Junior||1||70,000–100,000||35-50 Mb||35-50 Mb|
For more information take a look at Roche 454.