Tips for Pipetting Samples in qPCR Experiments

① Sample loading environment: qPCR experiments are highly sensitive, so the process of preparing the premix should be carried out in a clean bench to prevent contamination

② Preparation conditions: The premix reagents should be operated on ice and avoid direct exposure to strong light to reduce the possibility of template degradation, enzyme activity reduction, and fluorescence quenching during system preparation

③ Sample loading order: add large volume first, then small volume liquid; add NTC first, then template. Reduce operational errors and contamination risks

④　Sampling method:

➣ For small volume addition, the pipette can be used with the second gear for aspiration and the first gear for discharge (that is, press the bottom to aspirate, and do not press the bottom when discharging). If the tip is not replaced, the next hole will be aspirated at the first gear and punched at the second gear, which can improve the accuracy of sample addition and the repeatability of duplicate holes

➣ If the centrifuge tubes already have liquid, when adding a small volume, extend the gun below the liquid surface to avoid liquid residue in the gun tip

➣ When aspirating liquid, do not immerse the gun tip too deep below the liquid surface, and choose a low-adsorption gun tip to reduce errors caused by gun tip adsorption

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