The working principle of Opentrons Flex PCR Workstation is deeply rooted in the core principle of PCR (polymerase chain reaction), a molecular biology technology, and at the same time cleverly integrates Opentrons' cutting-edge technology and innovation achievements in the field of automated laboratory equipment research and development. Through a highly integrated automation system, this workstation not only accurately simulates the DNA replication process in nature, but also greatly improves the accuracy, repeatability and efficiency of experiments.
1. PCR reaction conditions PCR reaction conditions include temperature, time and number of cycles. The selection and optimization of these conditions are crucial to the success of the PCR reaction. 1. Temperature denaturation temperature: generally 90-95°C, used to completely denature the template DNA into a single strand. Annealing temperature: determined according to the Tm value of the primer, generally 40-60°C, used for the binding of the primer to the template DNA. Extension temperature: Generally 70-75°C, the DNA chain is extended under the action of Taq DNA polymerase. 2. Temporal denaturation time: Generally short, such as 1 minute, which is enough to completely denature the template DNA. Annealing time: Generally 30-60 seconds, which is enough for the primer to completely bind to the template DNA. Extension time: depends on the length of the fragment to be amplified, generally 1-15 minutes. 3. The number of cycles is generally 25-35. The higher the number of cycles, the greater the amount of amplification products, but it may also increase the production of non-specific products.
2. Preparation stage 1. Design primers: Design specific primers based on the target DNA sequence to ensure that the primers can efficiently bind to the template DNA and guide the DNA polymerase for amplification. 2. Prepare template DNA: Ensure that the purity and concentration of template DNA meet the requirements of the PCR reaction. Template DNA can be extracted from biological samples or DNA fragments from other sources. 3. Prepare reaction system: According to the needs of the PCR reaction, mix template DNA, primers, dNTPs (deoxynucleoside triphosphates), buffer, MgCl2 (if necessary), DNA polymerase, etc. in a certain proportion to form a PCR reaction system . This step needs to be performed in an environment free of nucleic acid contamination to prevent interference from foreign DNA. 4. Load the sample: Load the prepared PCR reaction system into the designated location of the Opentrons Flex PCR workstation, usually a PCR plate or PCR tube. Make sure the reaction volume and composition are consistent in each well or tube.
3. Program settings 1. Connect equipment: Connect the Opentrons Flex PCR workstation to the computer or other control equipment to ensure that the equipment can communicate normally. 2. Start the software: Open the control software or interface provided by Opentrons and prepare to set the PCR reaction program. 3. Create a new PCR program: Create a new PCR program in the software, and set parameters such as temperature, time, and cycle times for the three steps of denaturation, annealing, and extension. The setting of these parameters should be based on the characteristics of the target DNA sequence and the primers used. Opentrons Flex PCR workstations often support flexible programming capabilities that can be customized to suit experimental needs. 4. Save and upload the program: Save and upload the set PCR program to the Opentrons Flex PCR workstation. After ensuring that the procedure is correct, prepare to perform the PCR reaction.
4. Execute PCR reaction 1. Start the reaction: Click the "Start" or similar button in the software interface to start the PCR reaction. The Opentrons Flex PCR Workstation will automatically follow preset programs for temperature control and cycling. 2. Monitor the reaction: During the reaction process, the reaction status and progress can be monitored in real time through the software interface. Opentrons Flex PCR workstations usually have powerful monitoring functions that can display key information such as temperature curves and cycle times in real time. 3. End the reaction: When the PCR reaction reaches the preset number of cycles, the Opentrons Flex PCR workstation will automatically stop the reaction. At this point, the PCR plate or PCR tube can be removed for subsequent analysis.
5. Follow-up processing 1. Analyze PCR products: Analyze the specificity and yield of PCR products through agarose gel electrophoresis or other appropriate methods. Ensure that amplification products are of the expected size and quantity. 2. Data recording and sorting: record the results and data of the PCR reaction, and conduct necessary sorting and analysis. These data can be used for subsequent experimental verification or scientific research. 3. Cleaning and maintenance: After completing the PCR reaction, clean the Opentrons Flex PCR workstation and related equipment in time to prevent contamination and damage. At the same time, regularly check the performance and status of the equipment to ensure long-term stable operation of the equipment.
Please note that the above steps are general guidance based on general PCR reaction principles and functional features of the Opentrons Flex PCR Workstation. For specific operations, you should refer to the user manual of the Opentrons Flex PCR Workstation or contact the Opentrons technical support team for accurate guidance.
Related reading
Building a laboratory automation "PC"
White Paper: Opentrons High-Precision Pipettes
Handling White Paper: Temperature Control Module
Handling robot instrument service overview
The experienced service team and strong production support team provide customers with worry-free order services.
