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In the fields of biomedical research and clinical diagnosis, polymerase chain reaction (PCR) technology has become a key tool for amplifying specific DNA fragments due to its high efficiency and sensitivity. However, the cumbersome operating steps and possible manual errors in traditional PCR experiments often limit the accuracy and efficiency of the experiment. In order to overcome this challenge, Opentrons innovatively launched the OT-2 PCR workstation, which integrates two technologies: automated pipetting and precision temperature control, aiming to improve the accuracy and efficiency of experiments. The working principle and operating procedures of the Opentrons OT-2 PCR workstation will be explained in detail below.
Opentrons OT-2 PCR Workstation
1. Working principle 1. Principle of PCR technology: (1) PCR technology is a molecular biology technology that simulates the DNA replication process in vitro. Through the action of a series of enzymes, it can amplify and amplify specific DNA fragments. (2) Its specificity relies on oligonucleotide primers complementary to both ends of the target sequence, and the rapid amplification of DNA is completed through the three basic steps of denaturation-annealing-extension. 2. Automated pipetting platform: (1) Opentrons OT-2, as an automated pipetting platform, can accurately control the movement and operation of pipettes, thereby achieving precise preparation of PCR reaction solutions and accurate sample loading. (2) Through programming control, OT-2 can automatically complete the preparation of multiple PCR reactions, improving experimental efficiency and accuracy.
2. Work flow 1. Reagent preparation: (1) In the reagent storage and preparation area, use non-contaminated pipettes, pipette tips and containers to pack and prepare reagents. (2) Ensure that all PCR systems are stored at -20°C and the reagent components are completely melted before use to avoid affecting the concentration. 2. Sample preparation: (1) When receiving the sample, ensure the sealing and uniqueness of the sample number. (2) When performing sample operations in a biological safety cabinet, pay attention to the disinfection of the biological safety cabinet and the discharge of items. (3) Choose good quality Ep tubes for nucleic acid amplification reactions to ensure the stability and accuracy of the reaction system. 3. PCR reaction solution preparation: (1) On the Opentrons OT-2 automated pipetting platform, accurately prepare the PCR reaction solution according to experimental needs. (2) Premix the reagents except template DNA and distribute them into each PCR tube to ensure a consistent reaction system. 4. Add template DNA: (1) After thoroughly mixing the template DNA, use the Opentrons OT-2 automated pipetting platform to accurately add the sample into the PCR tube. (2) Pay attention to preventing cross-contamination and the generation of bubbles during the sample addition process. 5. Run the PCR program: (1) Place the PCR tube containing the reaction system into the PCR amplifier, and set appropriate temperature conditions and time parameters. (2) Start the PCR program and perform cyclic amplification of the three steps of denaturation, annealing and extension. (3) During the PCR process, the Opentrons OT-2 workstation can monitor the reaction progress and results in real time through the interface with the PCR amplification instrument. 6. Product analysis: (1) After the PCR program is completed, conduct agarose gel electrophoresis analysis or other appropriate analysis methods on the amplification product. (2) Judge the specificity and efficiency of PCR amplification based on the analysis results.
3. Precautions 1. During the entire PCR operation, experimental conditions must be strictly controlled to avoid contamination and errors. 2. Use high-quality reagents and consumables to ensure the accuracy and reliability of the experiment. 3. Regularly maintain and calibrate the Opentrons OT-2 workstation to ensure its normal operation and accuracy.
Opentrons OT-2 PCR workstation achieves efficient and accurate PCR operations by combining the core principles of PCR technology and the advantages of automated pipetting platforms. The workflow includes multiple steps such as reagent preparation, sample preparation, PCR reaction solution preparation, adding template DNA, running PCR programs and product analysis. Each step requires strict control of experimental conditions and operating specifications to ensure the accuracy and accuracy of experimental results. reliability.
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