Chief Technology Officer Stephen Turner of Pacific Biosciences will discuss Single Molecule Real-Time (SMRT) sequencing, due to be released commercially in 2010, at the 2009 Industrial Physics Forum, a component of the 51st Annual Meeting of American Association of Physicists in Medicine, which takes place from July 26 - 30 in Anaheim, California
A decade ago, it took Celera Genomics and the Human Genome Project years to sequence complete human genomes. In 2008, James Watson's entire genetic code was read by a new generation of technology in months. SMRT sequencing aims to eventually accomplish the same feat in minutes.
The method used in the Human Genome Project, Sanger sequencing, taps into the cell's natural machinery for replicating DNA. The enzyme DNA polymerase is used to copy strands of DNA, creating billions of fragments of varying length. Each fragment -- a chain of building blocks called nucleotides -- ends with a tiny fluorescent molecule that identifies only the last nucleotide in the chain. By lining these fragments up according to length, their glowing tips can be read off like letters on a page.
Instead of inspecting DNA copies after polymerase has done its work, SMRT sequencing watches the enzyme in real time as it races along and copies an individual strand stuck to the bottom of a tiny well. Every nucleotide used to make the copy is attached to its own fluorescent molecule that lights up when the nucleotide is incorporated. This light is spotted by a detector that identifies the color and the nucleotide -- A, C, G, or T.
By repeating this process simultaneously in many wells, the technology hopes to bring about a substantial boost in sequencing speed. "When we reach a million separate molecules that we're able to sequence at once … we'll be able to sequence the entire human genome in less than 15 minutes," said Turner.
The speed of the reaction is currently limited by the ability of the detector to keep up with the polymerase. The first commercial instrument will operate at three to five bases per second, and Turner reports that lab tests have achieved 10 bases per second. The polymerase has the potential to go much faster, up to hundreds of bases per second. "To push past 50 bases per second, we will need brighter fluorescent reporters or more sensitive detection," says Turner.
The device also has the potential to reduce the number of errors made in DNA sequencing. Current technologies achieve an accuracy of 99.9999 percent (three thousand errors in a genome of three billion base pairs). "For cancer, you need to be able to spot a single mutation in the genome," said Turner. Because the errors made by SMRT sequencing are random -- not systematically occurring at the same spot -- they are more likely to disappear as the procedure is repeated.
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ABOUT THE 2009 INDUSTRIAL PHYSICS FORUM
For each of the past 51 years, the Industrial Physics Forum has brought together industry, academic, and government leaders to examine applications of scientific research to emerging industrial R&D activities. This year's IPF is themed, "Frontiers in Quantitative Imaging for Cancer Detection and Treatment" and will be held in conjunction with the 51st Annual Meeting of the American Association of Physicists in Medicine (AAPM) on July 26 - 30, 2009 in Anaheim, CA. Embedded into the AAPM Scientific Program, the IPF sessions will be on Monday and Tuesday, July 27 - 28. During each IPF, a special session is dedicated to Frontiers in Physics, addressing the most exciting research going on today, regardless of field. In Anaheim, there will be speakers on next-generation DNA sequencers, on opto-genetics for brain imaging, and on how accelerator and particle physics enable some of the latest medical applications.
ABOUT MEDICAL PHYSICISTS
If you ever had a mammogram, ultrasound, X-ray, MRI, PET scan, or known someone treated for cancer, chances are reasonable that a medical physicist was working behind the scenes to make sure the imaging procedure was as effective as possible. Medical physicists help to develop new imaging techniques, improve existing ones, and assure the safety of radiation used in medical procedures in radiology, radiation oncology and nuclear medicine. They collaborate with radiation oncologists to design cancer treatment plans. They provide routine quality assurance and quality control on radiation equipment and procedures to ensure that cancer patients receive the prescribed dose of radiation to the correct location. They also contribute to the development of physics intensive therapeutic techniques, such as the stereotactic radiosurgery and prostate seed implants for cancer to name a few. The annual AAPM meeting is a great resource, providing guidance to physicists to implement the latest and greatest technology in a community hospital close to you.
The American Association of Physicists in Medicine (AAPM) is a scientific, educational, and professional organization of more than 6,000 medical physicists. Headquarters are located at the American Center for Physics in College Park, MD. Publications include a scientific journal (Medical Physics), technical reports, and symposium proceedings. See: www.aapm.org.
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