Polymerase chain reaction in polymeric microchips: DNA amplification in less than 240 seconds.
There is much interest in developing methods amenable to amplifying nucleic acids by the polymerase chain reaction (PCR) in small volumes in microfabricated devices. The use of infrared-mediated temperature control to accurately thermocycle microliter volumes in microchips fabricated from polyimide is demonstrated. Amplification of a 500-base-pair fragment of lambda-phage DNA was achieved in a 1.7-microl chamber containing a thermocouple that allowed for accurate control of temperature. While previous work showed that Taq polymerase was inactivated when in direct contact with the thermocouple, this was circumvented with the polyimide chip by the addition of polyethylene glycol as a buffer additive. This, consequently, allowed for adequate amounts of PCR product to be observed after only 15 cycles, with a total time for amplification of 240 s.
Department of Chemistry, University of Virginia, McCormick Road, Charlottesville, VA 22901, USA. (Work was carried out at Landers Lab, University of Pittsburgh, PA, USA.)
B C
BC
Giordano
J
J
Ferrance
S
S
Swedberg
A F
AF
Hühmer
J P
JP
Landers
D010582:Bacteriophage lambda / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D019075:Electrophoresis, Capillary / Q000379:methods; D007259:Infrared Rays; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011092:Polyethylene Glycols / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013696:Temperature
D011092:Polyethylene Glycols; D004247:DNA