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| Grant Number: | 5R44GM060113-03 |
| PI Name: | BONNER, ALEX G. |
| PI Email: | biolink@bellatlantic. net |
| PI Title: | PRESIDENT |
| Project Title: | HIGH-SPEED PCR USING A PRESSURIZED GAS THERMOCYCLER |
Abstract: DESCRIPTION (Applicant's abstract): In Phase I Research, a pressurized gas thermocycler was fabricated and successfully used to amplify gene fragments from human beta-globin (536 b.p.), Human Platelet Antigen (364 b.p.), bacteriophage lambda (333 b.p.), and a bacterial pathogen, E.coli 0157:H7 (91 b.p. and 85 b.p. amplicons). In our fastest experiments, a short 85 b.p. amplicon was amplified through 30 PCR cycles in 78 seconds. The speed and thermal control of this device were limited by its simple 3-valve configuration (hot gas valve, cold gas valve, throttle valve), flow conditioner, inadequate gas cooling, software, and lack of on-line DNA detection optics. In Phase II Research, improved high performance 4- and 5-valve thermocyclers will be fitted with on-line fluorescence detection optics. For short (less than 100 b.p.) amplicons, 30 cycles of amplification/detection will be carried out in less than 60 seconds using pressurized Helium gas. Longer amplicons (100 to 500 b.p.) will be amplified in 1 to 4 minutes using pressurized air. Pressurized gas thermocyclers will enable DNA-based diagnosis of heritable and infectious diseases on an unprecedented time scale. PROPOSED COMMERCIAL APPLICATION: The proposed method is ten times faster than the fastest commercially available hot-air PCR thermocyclers; and 100 times faster than commonly used heat block thermocyclers. Phase II Research will develop enabling technology for high-throughput-screening capabilities, which can be applied to diagnostics, drug discovery, and functional genomics related to heritable and infectious diseases. There is very large and growing market for high-speed PCR for medical/veterinary diagnostics, agribusiness, and biomedical research.
Thesaurus Terms:
biomedical equipment development, nucleic acid amplification technique, optics, polymerase chain reaction, technology /technique development, temperature
atmospheric pressure, computer program /software, computer system design /evaluation, fluorescence, genetic disorder diagnosis, helium, nucleic acid denaturation, nucleic acid hybridization
| Institution: | BIOLINK PARTNERS |
| 109 SCHOOL ST | |
| WATERTOWN, MA 02472 | |
| Fiscal Year: | 2002 |
| Department: | |
| Project Start: | 01-AUG-1999 |
| Project End: | 31-JUL-2003 |
| ICD: | NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES |
| IRG: | ZRG1 |