TH18650Y - Microfluidic series and microRNA analysis method using rolling circle amplification at a single temperature. - Google Patents

Abstract

------18/09/2563------(OCR) Microfluidic System and MicroRNA Analysis Procedure Using Rolling Circle Amplification at Single Temperature as described in this invention. A specific reagent set is used to analyze the target microRNA. The microfluidic system is optimized for stable temperature control. The RNA sequence amplification reaction within the microfluidic system occurs efficiently. Furthermore, the system is closed, preventing external contamination and allowing for use in remote areas without electricity, as the microfluidic system can be powered by batteries. The analysis of the target microRNA relies on measuring the fluorescence signal emitted from a single-layer graphene solution. ------------ Page 1 of 1 page. Abstract of the Invention. The microfluidic system and microRNA analysis procedure using rolling circle amplification at single temperature as described in this invention utilize a specific reagent set for the analysis of the target microRNA. The microfluidic system is optimized for stable temperature control. By maintaining a constant, controlled temperature, the RNA sequence multiplication reaction within the microfluid system occurs efficiently. Furthermore, it is a closed system, preventing external contamination, and can be used in remote areas without electricity because the microfluid system can be powered by batteries. The analysis of the target microRNA relies on measuring the fluorescent signal emitted from a single-layer graphene solution.

Claims (9)

18/09/2563 (OCR) 1. Microfluid array for the analysis of target microRNAs using the rolling circle amplification method at a single temperature. The microfluid array consists of: a. Channel 1 (111) for the initial fluid, channel 2 (112) for the detector, and channel 3 (113) for removing the fluid from the microfluid array; b. Heating section (121); c. Mixing section (122) with turbulent flow; and d. Detection section (131) using fluorescence principles. 2. Microfluid array according to claim 1, in which the heating section (121) is heated by a heating plate (400) and its temperature is predetermined to the temperature required for the rolling circle amplification method. The heating section (121) transfers heat to the fluid introduced into the microfluid array through channel 1 (111). 3. Microfluid array According to claim 1, the mixing section (122) is designed as a continuous S-shaped fluid channel to create turbulent flow, resulting in mixing of the fluid entering through channel 1 (111) after heating with the detector inserted through channel 2 (112).4. Microfluid set. According to claim 1, the detector section (131) is designed as a reservoir in the plane below the continuous S-shaped fluid channel, causing the mixed fluid to enter the reservoir in the detector section (131) for fluorescence detection.5. Microfluid set analysis procedure using single-temperature rolling circle amplification. According to claim 1, the procedure consists of: a. MicroRNA amplified using single-temperature rolling circle amplification in a microfluid set using an open-ended circular template DNA specific to the target microRNA. To obtain microRNA products, and b. Analysis of microRNA products using fluorescent agents and graphene materials. 6. MicroRNA analysis procedure according to claim 5, where the rolling circle amplification procedure is performed by mixing the target microRNA with reagent 1, which contains an open-ended circular DNA template specific to the target microRNA, to obtain a microRNA mixture. This mixture is then introduced into a microflow along with reagent 2, which contains deoxyribonucleoside triphosphate (dNTP), DNA polymerase enzyme, bovine serum albumin protein, and buffer solution. 7. MicroRNA analysis procedure according to claim 6, where the microRNA mixture and reagent 2 are mixed and introduced into a microflow and heated to amplify the target microRNA to obtain microRNA products. 8. MicroRNA analysis procedure. According to claim 5 or 7, where the microRNA product is mixed with a detector, fluorescently labeled DNA, in a single-layer graphene oxide solution for the analysis of the microRNA product.9. MicroRNA analysis procedure according to claim 5, where the analysis of the microRNA product is performed by measuring the fluorescent signal emitted from the single-layer graphene solution. ------------ ------05/04/2018------(OCR) Page 1 of 2 pages. Claim 1. A microfluid set for the analysis of target microRNA using the rolling circle amplification method at a single temperature. The microfluid set consists of: a. Channel 1 (111) for inserting the initial fluid, channel 2 (112) for inserting the detector, and channel 3 (113) for removing the fluid from the microfluid set; b. the heating section (121); c. the mixing section (122) with turbulent flow; and d. the detection section (131) using fluorescence principles. 2. Microfluid series under claim 1, in which the heating element (121) is heated by a heating plate (400) and its temperature is predetermined to the temperature required for the rolling circle amplification method. The heating element (121) is responsible for transferring heat to the fluid introduced into the microfluid series through channel 1 (111). 3. Microfluid sequence according to claim 1, in which the mixing section (122) is designed as a continuous S-shaped fluid channel to create turbulent flow resulting in mixing of the fluid entering through channel 1 (111) after heating with the detector inserted through channel 2 (112). 4. Microfluid series according to claim 1, in which the probe (131) is designed as a reservoir in a plane lower than a continuous S-shaped fluid channel, in which the mixed fluid is placed in the reservoir in the probe (131) for fluorescence measurement. 5. A procedure for the analysis of target microRNAs using a microfluidic system with single-temperature rolling circle amplification, as per claim 1, which comprises: a. The amplification of target microRNAs using single-temperature rolling circle amplification in a microfluidic system with an open-ended circular DNA template specific to the target microRNA to obtain microRNA products; and b. The analysis of the microRNA products using fluorescence and graphene materials. 6. MicroRNA analysis procedure as per claim 5, whereby the rolling circle amplification procedure is performed by mixing the target microRNA with reagent 1, which contains an open-ended circular DNA template specific to the target microRNA, to obtain a microRNA mixture solution. This solution is then placed in a microflow fluid along with reagent 2 (page 2 of 2), which contains deoxyribonucleoside triphosphate (dNTP), DNA polymerase enzyme, bovine serum albumin protein, and buffer solution. 7. MicroRNA analysis procedure under claim 6, in which a microRNA solution and reagent 2 are mixed and introduced into a microfluid stream and heated to induce the amplification of the target microRNA, yielding the microRNA product. 8. MicroRNA analysis procedure as per claim 5 or 7, where the microRNA product is mixed with a fluorescently labeled DNA detector in a single layer of graphene oxide solution for the analysis of the microRNA product. 9. MicroRNA analysis procedure according to claim 5, in which the analysis of microRNA products is performed by measuring the fluorescence signal emitted from a single-layer graphene solution. ------------ 1. Microfluid system for the analysis of target microRNAs by rolling circle amplification at a single temperature. The microfluid system consists of: a. Channel 1 (111) for the initial fluid, channel 2 (112) for the detector, and channel 3 (113) for removing the fluid from the microfluid system; b. Heating section (121); c. Mixing section (122) with turbulent flow; and d. Detection section (131) using fluorescence principles. 2. Microfluid system according to claim 1, in which the heating section (121) is heated by a heating plate (400) and its temperature is pre-controlled to a predetermined temperature for the rolling circle amplification method. 1. The heating section (121) is responsible for transferring heat to the fluid introduced into the microfluid system through channel 1 (111). 3. The microfluid system according to claim 1, where the mixing section (122) is designed as a continuous S-shaped fluid channel to create turbulent flow, resulting in mixing of the fluid introduced through channel 1 (111) after heating with the detector introduced through channel 2 (112). 4. The microfluid system according to claim 1, where the detection section (131) is designed as a reservoir below the continuous S-shaped fluid channel, causing the mixed fluid to be contained in the reservoir of the detection section (131) for fluorescence detection. 5. The procedure for analyzing the target microRNA using the rolling circle amplification method at a single temperature in the microfluid system. According to claim 1, the procedure consists of: a. The amplified target microRNA by rolling circle amplification at a single temperature in a microfluidic system using an open-ended circular DNA template specific to the target microRNA to obtain a microRNA product; and b. The analysis of the microRNA product using fluorescence and graphene materials. 6. The microRNA analysis procedure according to claim 5, where the aforementioned rolling circle amplification step is achieved by mixing the target microRNA with reagent 1, which contains an open-ended circular DNA template specific to the target microRNA, to obtain a microRNA mixture. This mixture is then introduced into a microfluidic system along with reagent 2, which contains deoxyribonucleoside triphosphate (dNTP), DNA polymerase enzyme, bovine serum albumin protein, and buffer solution. 7. The microRNA analysis procedure. According to claim 6, the microRNA mixture and reagent 2 are mixed and introduced into a microfluidic system and heated to induce the replication of the target microRNA, yielding microRNA products. 8. The microRNA analysis procedure according to claim 5 or 7, where the microRNA products are mixed with a fluorescently labeled DNA detector in a single-layer graphene oxide solution for microRNA product analysis. 9. The microRNA analysis procedure according to claim 5, where the microRNA products are analyzed by measuring the fluorescent signal emitted from a single-layer graphene solution.