454 Life Sciences

Source: Wikipedia, the free encyclopedia.

454 Life Sciences
Industry
Roche in 2007 and shut down by Roche in 2013 (production ceased mid-2016)
Headquarters
Branford, Connecticut
,
USA
ProductsGenome sequencers, reagents
ServicesSequencing of genetic samples

454 Life Sciences was a biotechnology company based in

Roche in 2007 and shut down by Roche in 2013 when its technology became noncompetitive, although production continued until mid-2016.[1]

History

454 Life Sciences was founded by Jonathan Rothberg[2] and was originally known as 454 Corporation, a subsidiary of CuraGen. For their method for low-cost gene sequencing, 454 Life Sciences was awarded the Wall Street Journal's Gold Medal for Innovation in the Biotech-Medical category in 2005.[3] The name 454 was the code name by which the project was referred to at CuraGen, and the numbers have no known special meaning.[4]

In November 2006, Rothberg, Michael Egholm, and colleagues at 454 published a cover article with

Neanderthal Genome Project to complete the sequence of the Neanderthal genome by 2009.[5]

In late March 2007,

Roche Diagnostics acquired 454 Life Sciences for US$154.9 million.[6] It remained a separate business unit.[7]
In October 2013, Roche announced that it would shut down 454, and stop supporting the platform by mid-2016.[8]

In May 2007, 454 published the results of Project "Jim": the sequencing of the genome of

James Watson, co-discoverer of the structure of DNA.[9][10]

Technology

454 Sequencing used a large-scale parallel pyrosequencing system capable of sequencing roughly 400-600 megabases of DNA per 10-hour run on the Genome Sequencer FLX with GS FLX Titanium series reagents.[11]

The system relied on fixing

ATP sulfurylase, and luciferase
was also packed into the well. The PicoTiterPlate was then placed into the GS FLX System for sequencing.

454 released the GS20

next-generation DNA sequencer on the market. In 2008, 454 Sequencing launched the GS FLX Titanium series reagents for use on the Genome Sequencer FLX instrument, with the ability to sequence 400-600 million base pairs per run with 400-500 base pair read lengths. In late 2009, 454 Life Sciences introduced the GS Junior System, a bench top version of the Genome Sequencer FLX System.[12]

DNA library preparation and emPCR

Genomic DNA was fractionated into smaller fragments (300-800 base pairs) and polished (made blunt at each end). Short adaptors were then ligated onto the ends of the fragments. These adaptors provided priming sequences for both amplification and sequencing of the sample-library fragments. One adaptor (Adaptor B) contained a 5'-biotin tag for immobilization of the DNA library onto streptavidin-coated beads. After nick repair, the non-biotinylated strand was released and used as a single-stranded template DNA (sstDNA) library. The sstDNA library was assessed for its quality, and the optimal amount (DNA copies per bead) needed for emPCR is determined by titration.[13]

The sstDNA library was immobilized onto beads. The beads containing a library fragment carried a single sstDNA molecule. The bead-bound library was emulsified with the amplification reagents in a water-in-oil mixture. Each bead was captured within its own microreactor where PCR amplification occurs. This resulted in bead-immobilized, clonally amplified DNA fragments.

Sequencing

Single-stranded template DNA library beads were added to the DNA Bead Incubation Mix (containing DNA polymerase) and were layered with Enzyme Beads (containing sulfurylase and luciferase) onto a PicoTiterPlate device. The device was centrifuged to deposit the beads into the wells. The layer of Enzyme Beads ensured that the DNA beads remained positioned in the wells during the sequencing reaction. The bead-deposition process was designed to maximize the number of wells that contain a single amplified library bead.

The loaded PicoTiterPlate device were placed into the Genome Sequencer FLX Instrument. The fluidics sub-system delivered sequencing reagents (containing buffers and nucleotides) across the wells of the plate. The four DNA

standard flowgram format
(SFF) files for downstream analysis.

See also

  • DNA Sequencing

Notes

  1. ^ Hollmer, Mark (October 17, 2013). "Roche to close 454 Life Sciences as it reduces gene sequencing focus". Fierce Biotech.
  2. ^ Park, Andrea (February 15, 2022). "Quantum-Si taps founder Rothberg, prolific medtech entrepreneur, as interim CEO". Fierce Biotech. Retrieved February 22, 2022.
  3. ^ Totty, Michael (October 24, 2005). "A Better Idea". The Wall Street Journal. Archived from the original on September 30, 2015. Retrieved March 13, 2017.
  4. ^ Pollack, Andrew (August 22, 2003). "Company Says It Mapped Genes of Virus in One Day". The New York Times.
  5. PMID 17108958
    .
  6. ^ "Roche - Roche acquires 454 Life Sciences to strengthen presence in ultra-fast gene sequencing". Archived from the original on August 29, 2012. Retrieved November 14, 2012.
  7. ^ "Roche snares 454's sequencing tech in buyout". FierceBiotech. March 28, 2007.
  8. ^ "Following Roche's Decision to Shut Down 454, Customers Make Plans to Move to Other Platforms". October 22, 2013.
  9. PMID 18421352
    .
  10. ^ "Project Jim: Watson's Personal Genome Goes Public". Archived from the original on November 21, 2008. Retrieved June 16, 2007.
  11. PMID 19246620
    .
  12. ^ "454 Life Sciences Unveils New Bench Top Sequencer, Significant Improvements to the Genome Sequencer FLX System Including 1,000 bp Reads for 2010" (Press release). 454 Life Sciences. November 19, 2009. Archived from the original on November 15, 2011. Retrieved November 19, 2009.
  13. PMID 20435675
    .
  14. PMID 19066560
    .
  15. PMID 16056220.{{cite journal}}: CS1 maint: numeric names: authors list (link
    )
Retrieved from "https://en.wikipedia.org/w/index.php?title=454_Life_Sciences&oldid=1166563308"