Automation micro-flow workstation and method for carrying out solid phase PCR reaction

Abstract

The invention discloses an automation micro-flow workstation and a method for carrying out a solid phase PCR reaction. The method refers to a method for realizing the solid phase PCR reaction based on a micro-fluidic chip. The workstation comprises at least one micro-fluidic chip, a fluid path system, a temperature control device and a control system, wherein each chip comprises one or more independent reaction channels; each reaction channel comprises an inlet, an outlet, one or more reaction chambers and / or flow channels connected in series or in parallel; an oligonucleotide sequence is anchored to the bottom surface of each reaction chamber so as to perform solid phase PCR amplification and / or solid phase single base extension; the fluid path system controls to introduce different reaction reagent solutions so as to flow to corresponding reaction chambers; the temperature control device is used for controlling and regulating the temperature in the reaction chambers; and the control system is connected with the fluid path system and the temperature control device and is used for controlling the solid phase PCR amplification and / or solid phase single base extension performed in the reaction chambers of the micro-fluidic chips in the fluid path system and the temperature control device, and the reactions comprise a denaturation-elution reaction and an elution-denaturation reaction. The workstation has the advantages of one-stop automation, high flux, low cost, convenience in operation, rapidness, high efficiency and high accuracy.

Description

technical field [0001] The present invention relates to the field of genetic engineering, in particular to a microfluidic chip-based solid-phase PCR & single base extension reaction, docking nucleic acid mass spectrometry detection, mainly used in single nucleotide polymorphism detection, especially a method suitable for a Station automation, multi-sample, high-throughput, fast response, and low-cost SNP detection sample pretreatment can be directly connected with nucleic acid mass spectrometers such as MALDI-TOF-MS for SNP site detection. Background technique [0002] Single nucleotide polymorphisms (Single Nucleotide Polymorphisms, SNPs) refer to changes or mutations in DNA sequences caused by changes in single bases A, T, C, and G at specific nucleotide positions in the individual genome. Such mutations include Single base conversion, transversion, insertion or deletion, etc. Strictly speaking, only when the allele frequency is greater than or equal to 1%, it can be call...

AI Technical Summary

Problems solved by technology

The workstation is based on the bottom surface of the microfluidic chip channel as a solid-phase substrate to anchor and connect reverse primers, and perform solid-phase PCR amplification reactions, denaturation-elution reactions, one-step single-base extension reactions, and elution-denaturation reactions: to overcome In the existing mass spectrometry detection SNP sample processing process, it is necessary to perform two multi-cycle PCR amplification reactions and SAP digestion processing on the sample, resulting in high cost, time-consuming, laborious and other defects; with the help of liquid collection system, temperature control device and control The system is equal to the integrated microfluidic workstation to realize the one-stop automatic operation of the reaction; at the same time, the single-use chip avoids cross-contamination between samples, and can use different types of chips and different oligonucleotides according to different SNP sites and detection requirements acid sequence; after processing the sample at the workstation, it can be directly connected to the nucleic acid mass spectrometer; the workstation has the advantages of one-stop automation, high throughput, low cost, convenient operation, fast and efficient, and high accuracy

Images