System and methods for monitoring the amplification and dissociation behavior of DNA molecules

摘要

The present invention relates to systems and methods for monitoring the amplification of DNA molecules and the dissociation behavior of the DNA molecules. A method according to one embodiment of the invention may include the steps of: forcing a sample of a solution containing real-time PCR reagents to move though a channel; and while the sample is moving through an analysis region of the channel, performing the steps of: (a) cycling the temperature of the sample until the occurrence of a predetermined event; (b) after performing step (a), causing the sample's temperature to gradually increase from a first temperature to a second temperature; and (c) while the step of gradually increasing the sample's temperature is performed, using an image sensor to monitor emissions from the sample.

主权项

1. A method, comprising:
(a) introducing into a microchannel at least one bolus containing nucleic acid; (b) forcing the bolus to move through the microchannel; (c) while the bolus is moving through the microchannel, (i) causing a thermal generating apparatus to cycle the temperature of the bolus to amplify nucleic acid contained in the bolus, wherein the thermal generating apparatus sequentially controls temperature during amplification and thermal denaturation, the thermal denaturation being performed after the amplification is completed and (ii) at least once per temperature cycle determining whether a predetermined emission intensity threshold has been met, wherein the step of determining whether the predetermined emission intensity threshold has been met comprises using an image sensor to capture an image of the bolus and processing the captured image data to determine the intensity of light emitted from the bolus; (d) in response to determining that the predetermined emission intensity threshold has not been met, then repeating step (c); and (e) in response to determining that the predetermined emission intensity threshold has been met, then, causing said thermal generating apparatus to cease the temperature cycling and causing said thermal generating apparatus to gradually increase the temperature of the bolus, while the bolus is still moving through the microchannel, causing thermal denaturation of dsDNA within the bolus to transition to ssDNA and using the image sensor to capture images of the bolus.