DNA/RNA synthesizerThe usage method of (taking DNA as an example) includes multiple steps such as reagent preparation, sequence input, parameter setting, synthesis column inspection, pipeline cleaning, synthesis process monitoring, and product processing. The following is a detailed introduction:
Reagent preparation:
Carefully check the amount of reagents in all reagent bottles on the synthesizer, replace them if necessary, and pay special attention to solvents with large amounts such as acetonitrile.
Ensure that the reagent bottle group and base bottle group are well sealed, maintain a certain pressure, and prevent external air from entering and affecting the synthesis effect.
Sequence input:
The DNA sequence to be synthesized can be input into a synthesizer using computer-aided design software, taking into account factors such as codon preference, secondary structure, and restriction enzyme sites.
Check if the selected synthesis steps are correct and ensure that the synthesis process follows the preset procedure.
Parameter settings:
Choose the termination method. The original state of the instrument is usually TrityloOfAuto. If you plan to connect purified oligonucleotides with 5 '- DMT groups, switch it to the TrityloOnAuto termination state.
Under the Username of each column monitor, enter the desired save name for easy management and tracking in the future.
Use code instead of corresponding DNA sequence to label each collection bottle for easy identification and classification.
Composite column inspection:
Run the StartColumn step to check the position of each column, ensuring that the synthesis column on the synthesizer is in the correct position to avoid synthesis failure due to positional deviation.
Check if the solvent residue (waste liquid bottle) connected to the synthesizer is full to prevent liquid overflow caused by overfilling of the waste liquid bottle, which may affect the synthesis environment.
Pipeline cleaning:
If the branch collector is used to collect the effluent from each DMT deprotection step, ensure that the test tube is thoroughly cleaned, and open the branch collector to ensure that the DMT waste liquid pipeline leads to the branch collector.
Start the cycle, run the program to clean the reagent pipeline, remove the original reagents, and prevent old reagents from interfering with the new synthesis reaction.
Synthesis process monitoring:
During the synthesis process, the total conductivity of DMT cations released in each DNA synthesis cycle can be measured using a conductivity flow cell to monitor the progress of the synthesis reaction.
Regularly check the operating status of the synthesizer to ensure that all parameters (such as temperature, pressure, time, etc.) are within the normal range.
DNA/RNA synthesizerSynthesis completion and product processing:
After synthesis, the vector is removed for cutting and deprotection. Generally, DNA fragments within 40bp require at least 2 hours, and longer chains require longer time.
Adopt appropriate purification methods (such as OPC purification, PAGE purification, desalination purification, HPLC purification, etc.) to remove short fragments and salt ions and improve product purity.
Perform sequencing validation on the purified DNA to ensure its sequence matches expectations.