Power source-free and valve-free type single molecule detection chip and applications thereof
A single-molecule detection, valveless technology, used in laboratory containers, microbial determination/inspection, bioreactor/fermenter combination, etc. Many other problems, to achieve the effect of improving the speed of the experiment, reducing the consumption and reducing the cost of the experiment
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Embodiment 1
[0056] see figure 1 , a microfluidic chip device for nucleic acid amplification that does not require an external drive system, does not require valves, has a simple manufacturing process and is easy to operate. The device consists of a reaction layer 1, a modification layer 2, a sealing layer 3, a main channel 4, a reaction chamber 5, a sample inlet 6, a sample outlet 7, and a heat-resistant tape 8. The reaction layer 1 is composed of a main channel 4 in series with reaction chambers. 5, the reaction chamber 5 is located above the main channel 4, the sample inlet 6 and the sample outlet 7 are located at both ends of the main channel 4, the modification layer 2 is connected to the sealing layer 3 in advance, and the reaction layer 1 is connected to the sealing layer through the modification layer 2. 3 After sealing, the microfluidic chip assembly is formed. A heat-resistant adhesive tape 8 is pasted on the upper surface of the reaction layer 1 .
[0057] The reaction layer 1...
Embodiment 2
[0068] see figure 2 , a microfluidic chip device for nucleic acid amplification that does not require an external drive system, does not require valves, has a simple manufacturing process and is easy to operate. The device consists of a reaction layer 1, a modification layer 2, a sealing layer 3, a main channel 4, a reaction chamber 5, a sample inlet 6, a sample outlet 7, and a heat-resistant tape 8. The reaction layer 1 is composed of a main channel 4 in series with reaction chambers. 5, the reaction chamber 5 is located below the main channel 4, the sample inlet 6 and the sample outlet 7 are located at both ends of the main channel 4, the modification layer 2 is connected to the sealing layer 3 in advance, and the reaction layer 1 is connected to the sealing layer through the modification layer 2. 3 After sealing, the microfluidic chip assembly is formed. A heat-resistant adhesive tape 8 is pasted on the upper surface of the reaction layer 1 . The reaction layer 1 is made...
Embodiment 3
[0078] Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) on a valve-free microfluidic PDMS chip without power source.
[0079] In this example, the device in Example 1 is used to inject samples, and the subsequent real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) is performed. Specific steps:
[0080] 1. See image 3 Channel structures are designed and masked.
[0081] 2. The reaction layer 1 is made of polydimethylsiloxane (PDMS), and a substrate with a microchannel structure produced by multi-layer soft lithography technology is used as a mold, and PDMS with gas permeability is poured on the mold and solidified. The reaction chamber 5 is a square with a side length of 100 μm, 200 μm or 300 μm, and a depth of 50 μm, 100 μm, 200 μm, or 300 μm. According to different widths and mold heights, the volume of the reaction chamber 5 can be 500 pL, 1 nL, 2 nL, 4.5 nL, 8 nL, 9 nL, 18 nL; the reaction chamber 5 can also be penta...
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