The preparation and loading of PCR reaction samples is a very important operation throughout the PCR reaction. Manual loading is difficult to ensure the accuracy and accuracy of each hole. Only automated sample loading stations ensure the reliability of micro-dosing.

Twenty years ago, Kary Mullis invented the polymerase chain reaction (PCR), which caused major changes in many fields of life science research and medicine. Since then, this new technology of DNA replication has continued to evolve. The latest example is the application of the detection of severe acute respiratory syndrome (SARS). Coronavirus pathogens were detected without errors by polymerase chain reaction. PCR reaction characteristics require special conditions and techniques. In practice, PCR can amplify 10 to 100 DNA molecules. This highly sensitive and specific PCR has high application value. The preparation and loading of PCR reaction samples is a very important operation throughout the PCR reaction. Manual loading is difficult to ensure the accuracy and accuracy of each hole. In particular, the distance between the holes of the 384-well plate is only 4.5 mm, so the proficiency of the sampler is very high. Only automated sample loading stations ensure the reliability of such micro-loading.

Say goodbye to the tedious and heavy pipetting work

Many molecular biology methods have been automated, and the field of automation is still expanding, and has begun to replace traditional manual methods.
Eppendorf's pipetting station epMotion® 5070 is used for real-time quantitative PCR (qPCR) systems compared to manual method qPCR systems. The pipetting station epMotion 5070 involves operations including standard gradient dilution, a series of transfer liquids, and loading into the reaction wells.

To amplify specific target genes and detect amplification signals, Eppendorf's RealMasterMix Probe reagent, the only optimized qPCR for hydrolysis probes, is characterized by its proprietary HotMaster®. Taq DNA polymerase is suitable for the establishment of a PCR reaction at room temperature.

Two qPCR platforms were used: ABI Prism 7000SD (Applied Biosystem, Foster City, CA, USA) and Bio-Rad iCycler iQ (Bio-Rad, Hercules, CA, USA).

In short, the real-time qPCR system, which is part of the Eppendorf automated pipetting station epMotion® 5070, reduces the number of common defects in manual handling compared to manual method PCR, while improving repeatability and accuracy, as well as overall pollution reduction. possibility.

Easy to complete the sample loading work

Workers who often use 96-well or 384-well plates for PCR experiments, or who often process enzyme-labeled plates, must realize that such tests require a high concentration of attention. It’s been a long time and I feel exhausted. Some mistakes often occur when operating such tests. For example, after changing the nozzle, have you forgotten that the primer in the 23rd column has been added? Never know whether it starts with column 22 or column 23? With the epMotion 5070 small autosampler station, there are no more problems like this. In just 7 minutes, reagents required for the experiment, such as HotMasterMix, molecular biology grade purified water, or 96 different samples, can be added to the 96-well plate. If it is a 384-well plate, it may take 21 minutes, because the epMotion 5070 is a small sample loading station. If the same sample is divided into 96 wells, the whole process takes only 2 minutes!

Perfect combination

The automatic sample loading station can handle all the repeated sample loading work, which is without a doubt. If you work with other Eppendorf products such as reagents and consumables, you can provide a perfect experimental condition for PCR experiments. For example, Eppendorf's HotMasterMix ready-to-use kit is used for hot-start PCR reactions, which is ideal for automated sample processing stations. Because the HotMaster hot start enzyme can be stored in a normal refrigerator without prior thawing, and the kit can be placed in the bench for a long time without cooling. Its hot start performance characteristics prevent pre-reaction of the enzyme with other components. The reason is that the HotMaster kit has a patented HotStart / Cold Stop technology that achieves a hot start/cold stop effect throughout the PCR cycle. Different from other traditional hot start kits. Other brands of hot-start enzymes, after activation at 95 ° C, do not restore the function of Taq polymerase at low temperatures. Moreover, the use of the kit eliminates the need to explore the appropriate magnesium ion concentration, because MasterMix's buffer system has the function of self-regulating magnesium ion concentration and can be adapted to various PCR reaction conditions.

The 96,384 twin.tec PCR plate is especially suitable for automated sample loading systems and is SDS compliant. The edge of the twin.tec PCR plate is made of polycarbonate and the thin-walled walls are made of polypropylene. This material and manufacturing process can withstand high temperature and mechanical pressure, and the experimental deviation is small.

Automatic loading requires a high demand for the material and shape of the nozzle. The epMotion-specific nozzle epT.IPS is placed in a non-contaminating nozzle box and is available in two different purity nozzle sets of premium grade and PCR clean grade. According to the experimental needs, you can also choose a filter with a filter element and no filter element.

No longer boring

The most fortunate thing is that you don't have to worry about repeatability, and you don't have to worry about forgetting the order of loading. From then on, this tedious, but repetitive work must be handed over to epMotion, which is not only highly automated, but also reliable.

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