Convective polymerase chain reaction apparatus and optical detecting method thereof

Abstract

The invention discloses a convective polymerase chain reaction apparatus and an optical detecting method thereof. The convective polymerase chain reaction apparatus includes a tube, a temperature control unit, at least one light source and a sensor. The tube includes a cavity used to contain a reaction solution. The reaction solution has a liquid level measured from a bottom of the cavity to a topsurface of the reaction solution. The temperature control unit is disposed adjacent to the tube for controlling the temperature of the reaction solution. The at least one light source provides a light beam passing through an incident portion of the tube to excite the reaction solution emitting a fluorescent light. The incident portion is located at a height greater than 1 / 2 of a liquid level. Thesensor is adjacent to the tube for detecting the excited fluorescence. The light beam has an incident direction forming a non-straight angle with a long axis of the tube.

Description

technical field [0001] The invention relates to a polymerase chain reaction (Polymerase Chain reaction; PCR) device and a detection method thereof. In particular, it relates to a convective polymerase chain reaction (cPCR) device and an optical detection method thereof. Background technique [0002] Polymerase chain reaction is a molecular biology technique for amplifying specific deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) fragments. A typical polymerase chain reaction device uses repeated heating and cooling of a thermocycler to perform repeated thermal cycle steps, and the heat energy is gradually conducted (thermal conduction) into the reaction test tube through the metal block, so that DNA or Ribonucleic acid fragments are subjected to three different temperature cycles of denaturation (95°C), adhesion (45°C-65°C) and extension (72°C) at different temperatures. However, the traditional way of using multi-level indirect heating for thermal cycle steps is not ...

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Problems solved by technology

[0004] However, since the reaction liquid contains biological samples (such as the whole blood of the organism), its composition is quite complicated

And during the reaction operation, most of the contents of the reaction liquid (such as protein, blood cells, etc.) will be deposited at the bottom of the capillary tube, causing interference and obstruction on the excitation light path, making it impossible for the light to pass through the bottom of the tube to excite the fluorescent reagent in the tube. The fluorescent signal cannot be sensed by the detector on the side of the tube, which seriously affects the control quality of the polymerase chain reaction

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