The OT-2 automated pipetting platform is one of the main products of Opentrons. When conducting protein extraction experiments, its automated pipetting platform and supporting modules are mainly used, such as the OT-2 automated pipetting platform and magnetic bead purification module. , to assist in completing the entire extraction process. Although Opentrons itself does not directly provide a specific "protein extractor", its automated solutions can greatly improve the efficiency and accuracy of protein extraction.
1. Equipment for protein extraction. Specifically, the application of Opentrons in protein extraction may include the following instruments and modules: 1. OT-2 automated pipetting platform: This is one of the core products of Opentrons, which can accurately carry out liquid Transfer, mix, distribute and other operations. During the protein extraction process, OT-2 can automatically draw reagents such as lysis buffer from the reagent bottle and distribute them into centrifuge tubes containing samples, thereby reducing errors caused by manual operations. 2. Magnetic bead purification module: Opentrons also provides a purification module based on the principle of magnetic bead extraction, which can be used for protein purification and concentration. Magnetic beads can bind to specific proteins and then separate the magnetic beads from other components in the solution through the action of the magnetic field, thereby purifying the protein. Opentrons' magnetic bead purification module can be used in conjunction with the OT-2 automated pipetting platform to realize automated protein purification processes. 3. Other supporting modules: In addition to the above-mentioned pipetting platform and purification module, Opentrons also provides supporting equipment such as thermal shakers and temperature control modules. These equipment can provide necessary temperature control, shaking and mixing during the protein extraction process. operation to optimize the extraction effect.
2. Protein extraction steps of OT-2 automated pipetting platform 1. Sample preparation Cell collection: For suspended cells, collect the cells by centrifugation; for adherent cells, use trypsin and other treatments to detach the cells before collecting. Wash: Use an appropriate buffer (such as PBS) to wash the cells to remove residual media and impurities. 2. Lysis and mixing. Preparation of lysis solution: Prepare lysis solution containing protease inhibitors (such as RIPA lysis solution) on the OT-2 platform. Lysis operation: Automatically distribute the lysis solution into centrifuge tubes containing cells, and mix thoroughly through the mixing function of OT-2 to ensure sufficient cell lysis. 3. Treatment of lysate products: Ice bath lysis: Place the centrifuge tube on ice and use the oscillation function of OT-2 to oscillate for a certain period of time to promote cell lysis. Centrifugation: Although OT-2 itself does not directly perform centrifugation operations, it can be used with a centrifuge. The lysed mixture is centrifuged to separate the supernatant (i.e., protein solution). 4. Protein purification (optional) If you need to further purify the protein, you can use OT-2 with the magnetic bead purification module. Mix the magnetic beads with the protein solution, bind the magnetic beads to the protein through the action of a magnetic field, then wash to remove impurities, and finally separate the magnetic beads and the protein through a magnetic field to obtain purified protein. 5. Protein quantification and analysis Protein quantification: Use an appropriate protein quantification method (such as BCA method) to quantify the extracted protein. Analysis: According to the experimental requirements, the extracted proteins are subjected to subsequent analysis and detection, such as SDS-PAGE electrophoresis, Western Blot, etc.
3. Precautions 1. During the entire experimental process, it is necessary to ensure that all reagents and consumables are sterile and pure to avoid contamination of the experimental results. 2. Experimental procedures and conditions should be adjusted and optimized according to specific experimental needs and sample characteristics. 3. The operation of the OT-2 automated pipetting platform requires prior programming and settings through software such as Protocol Designer to ensure the accuracy and repeatability of the experiment. 4. When using OT-2 for protein extraction, it is recommended to refer to the detailed experimental steps and precautions on the Opentrons official website or related literature.
The above steps are a general description of protein extraction based on the OT-2 automated pipetting platform. The specific experimental steps and conditions may vary depending on experimental needs and experimental conditions. In actual operation, it is recommended to adjust and optimize according to specific conditions.
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