The Corbett Research RG 3000 is a reliable thermal cycler, allowing researchers to perform PCR reactions quickly while also giving them the ability to monitor the reaction in real-time. The cycler holds either 36-0.2 ml or 72-0.1 ml standard PCR tubes and it is an open chemistry platform. The open chemistry platform uses multiple excitation sources (470, 530, 585 and 625 nm LED high power diodes) combined with several detection filters (510, 555 and 610 nm bandpass and 665, 570 and 610 nm high-pass) to detect virtually every known fluorophore (Sybr-Green I, FAM, TET, JOE, VIC, HEX, ROX, TAMRA, CY3, CY3.5, CY5, CY5.5, Oregon Green, CAL Red, Red 640 and Texas Red). Also, most DNA amplification enzymes/ buffers can be used. In our lab, we have used three different enzymes without problems. The system comes with data analysis software that is constantly upgraded and very easy to use.
I have used the RG3000 for quantitative RT-PCR with Sybr-Green I and FAM labeled LUXTM primers with subsequent melt curve analysis. Both dyes worked perfectly and the RG software makes is easy to design the templates and quantify the products. I have not used the cycler for other possible uses, such as allelic discrimination, SNP detection or analyzing multiple targets in the same reaction using differentially labeled hybridization probes. The optical denaturation is a newly introduced feature, giving you the possibility to monitor fluorescence during denaturation. Once this is done, samples will have to be heated only to the necessary temperature to reach total denaturation. This can shorten the run times up to 25%.
Because Sybr-Green I is an inexpensive dye, we were able to test and establish primers for around 30 different targets in one year. When trying a new primer set, the use of normal PCR tubes makes it easy to visualize products on agarose gels for verification (if the melt curve analysis is not sufficient). As usual with PCR, accurate quantification depends on accurate pipetting, because even small errors are amplified during the process. When performing time course experiments with many samples, one sometimes wished to have bigger rotors with maybe up to 96 samples, because the offered rotor with 72 samples was sometimes simply to small, forcing You to perform two runs of the same target. My only experience with the product support is regarding the download of regular published software updates from the Corbett Research homepage. In our lab we never had any problems with this cycler, so I can’t really comment on the support regarding other issues.
The main advantage of the RG3000 is the flexibility and the low running cost of the cycler. As stated above, more or less every dye and amplification enzyme can be used. Also, the reaction vessels are standard PCR tubes and no expensive kits have to be used. Once a run is established, you might find yourself doing runs that are not always necessary, just because it is so easy and effective.
Carl Gebhardt, MD
University of Leipzig