PCR amplification process of Opentrons Flex PCR workstation

In the vast field of biological sciences, polymerase chain reaction (PCR) has always been an important tool in molecular biology experiments. This technology can quickly replicate minute amounts of DNA samples, providing a solid foundation for subsequent research. The Opentrons Flex PCR workstation, which combines advanced robotic technology and precise temperature control system, makes PCR amplification more stable and reliable. Whether they are just beginning students or experienced researchers, this platform can help them easily achieve efficient and accurate PCR amplification.

Opentrons Flex PCR Workstation

1. Experimental preparation stage 1. Design primers: Use professional primer design software to design appropriate primer pairs based on the target DNA sequence. Primer design needs to consider factors such as specificity, length, GC content, and annealing temperature. 2. Prepare template DNA: Extract the required DNA template and determine its concentration and purity. Ensure that the concentration and purity of the template DNA meet the requirements of the PCR reaction. 3. Prepare the PCR reaction system: According to the experimental needs, prepare the PCR reaction mixture, including DNA template, primers, dNTPs (deoxynucleoside triphosphates), PCR buffer, MgCl₂ (if necessary), Taq DNA polymerase, etc. Mix all ingredients in a certain proportion and distribute into PCR tubes.

2. Sample loading and instrument setting stage 1. Preheat the Opentrons Flex PCR workstation: Open the workstation and set the preheating temperature according to the experimental requirements, usually 94-96°C. 2. Load the sample: Add the prepared PCR reaction mixture into the PCR tube. Place the PCR tubes in the sample rack of the Opentrons Flex PCR Workstation. 3. Set PCR cycle parameters: On the control interface of the workstation, set the cycle parameters of PCR amplification, including the temperature and time of denaturation, annealing and extension, as well as the number of cycles. Parameter settings need to be optimized based on specific experimental requirements and the characteristics of the target DNA sequence.

3. PCR amplification stage 1. Denaturation step: Heat the reactor to about 93-98°C and keep it for a certain time (such as 20-30 seconds) to dissociate the DNA double strands into single strands. 2. Annealing step: Lower the temperature to about 50-65°C and maintain it for a certain period of time (such as 20-40 seconds) to pair the primer with the complementary sequence of the template DNA single strand. 3. Extension step: Raise the temperature to the optimal temperature of DNA polymerase (usually 72°C) and maintain it for a certain period of time (depending on the length of the target DNA fragment and the ability of the DNA polymerase), so that the DNA polymerase uses dNTPs as the raw materials to synthesize new DNA strands. 4. Cyclic amplification: Repeat the above denaturation, annealing and extension steps to form a cycle of PCR amplification. Typically a PCR process uses 25-35 cycles, with each cycle doubling the number of target DNA fragments.

4. End of amplification and result analysis stage 1. Final extension: After all cycles are completed, an additional extension step is usually performed at 72°C to ensure that all unfinished DNA strands are fully extended. 2. Cool down and remove samples: After amplification, the Opentrons Flex PCR workstation will automatically cool down to 4°C and maintain it. At this time, the sample can be removed. 3. Result analysis: Analyze the PCR products through gel electrophoresis, real-time fluorescence quantitative PCR and other methods. Observe the DNA bands in the electrophoresis pattern to determine whether the PCR amplification is successful and the specificity and concentration of the amplified product.

5. Post-experiment processing and instrument maintenance stage 1. Clean the work area: After the experiment, clean the work area in time to avoid cross-contamination. 2. Data recording: Record experimental results, including amplification curves, electrophoresis patterns, etc., to provide reference for subsequent experiments. 3. Instrument maintenance: Regularly maintain and calibrate the Opentrons Flex PCR workstation to ensure the accuracy and stability of the instrument.

The PCR amplification process of the Opentrons Flex PCR workstation includes multiple stages such as experimental preparation, sample loading and instrument setup, PCR amplification, amplification completion and result analysis, as well as experimental post-processing and instrument maintenance. Each stage requires careful operation to ensure the accuracy and reliability of PCR amplification.

Related reading recommendations

Opentrons New York ISO 9001 certification

How the COVID-19 epidemic has revolutionized the life sciences

Handling White Paper: HEPA Module

Advantages of fully automatic pipetting workstations for culture medium preparation

Tip specifications of OT-2 nucleic acid extraction workstation

Opentrons Flex NGS Library Preparation Workstation Functions