Real-time PCR in micro-channels
First Claim
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1. A method of performing real-time PCR comprising the steps of:
- a) continuously moving a bolus of test solution containing real-time PCR reagents in a channel;
b) moving a carrier fluid in said channel, sequentially alternating with a test bolus;
c) cycling the temperature in a defined section of the channel in order to achieve PCR, wherein said defined section of the channel has a uniform temperature, which temperature cycles in time; and
d) measuring the intensity of the fluorescent signal at a plurality of locations along said defined section of said channel.
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Abstract
The present invention relates to methods for amplifying nucleic acids in micro-channels. More specifically, the present invention relates to methods for performing a real-time polymerase chain reaction (PCR) in a continuous-flow microfluidic system and to methods for monitoring real-time PCR in such systems.
38 Citations
32 Claims
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1. A method of performing real-time PCR comprising the steps of:
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a) continuously moving a bolus of test solution containing real-time PCR reagents in a channel; b) moving a carrier fluid in said channel, sequentially alternating with a test bolus; c) cycling the temperature in a defined section of the channel in order to achieve PCR, wherein said defined section of the channel has a uniform temperature, which temperature cycles in time; and d) measuring the intensity of the fluorescent signal at a plurality of locations along said defined section of said channel. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A method for monitoring the progress of a polymerase chain reaction in a channel, comprising the steps of:
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a) moving a bolus of test solution containing real-time PCR reagents in a channel; b) moving a carrier fluid in said channel, sequentially alternating with a test bolus; c) cycling the temperature in a defined section of the channel in order to achieve PCR, wherein said defined section of the channel has a uniform temperature, which temperature cycles in time; and d) capturing an image of a reaction-dependent fluorescence signal along a section of the channel; e) measuring the average flow speed in the channel; and f) relating position of the test bolus to the number of temperature cycles experienced by a test bolus from the average flow speed. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32)
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Specification