MasterCycler RealPlex⁴ From Eppendorf

This is Eppendorf’s big-brother qPCR platform, which in many ways is similar to the RealPlex². Like the RealPlex², this machine is small, solid and very quiet. It has a small footprint (40cm x 24cm x 40cm), which should appeal to those working in small or over-crowded laboratories. The RealPlex⁴ carries a fast 96-well heating block, which cuts down assay run-time. The system can also generate a thermal gradient from left to right, which is easy to set up and aids PCR optimization greatly. Depending on the complexity of the PCR assay, the RealPlex⁴ performs four to five complete runs per day, including set-up and report generation.

The RealPlex⁴ has four color filters, enabling fluorescence monitoring at the following wavelengths: 520, 550, 580 & 620 nm. This means that it is immediately compatible with the majority of reporting dyes. Importantly, for people who routinely use TaqMan® Mastermix from Applied Biosystems, the RealPlex⁴ can detect ROX passive dye, and easily normalize base-line fluorescence levels, minimizing experimental variation between multiplets, and giving more accurate results. This can certainly be useful in highly sensitive quantitative assays.

Both RealPlex machines use the same software interface, and it’s a great one, which is a piece of cake to use. The software does a great job of saving and archiving all of your data, making it almost impossible to lose or overwrite anything. Also, the user has complete freedom to set-up a 96-well plate in any way (unlike other, more prescriptive machines that try to do the thinking for you).

Again, the RealPlex⁴ uses the same fixed 96 LED method for generating excitation light. Hence, there are no moving parts, making the fluorescence monitoring very quiet. In contrast, other machines use a moving detection head, which is noisy and obviously slower.

I would definitely recommend this instrument for any research lab. It’s flexible enough for any real-time assay, and can deal with multiplexing and SNPs easily. I also think it represents good value for money (in terms of its features, and quick cycling time). I also think that Eppendorf’s after-sales service is pretty good, which is an important consideration, especially if you’re buying a suite of machines. At the moment, it is not possible to export numerical fluorescence data from the software. This means that you can only use the fluorescence plots that the machine generates automatically. This is not really a big problem, because you can control colors and data-labels easily. However, I do think the instrument would be even more useful if it could export all fluorescence data direct to a spreadsheet. I have been told that this may be addressed in future software updates.