7300 Real-Time PCR System From Applied Biosystems

Polymerase Chain Reaction (PCR), based upon my opinion, is perhaps the single technique that has most revolutionized molecular biology. The recent advances in PCR and especially the development of quantitative real-time PCR (qPCR, also known as real-time PCR) make that methodology a staple of almost any biological laboratory, with myriads of applications in almost any bioscience discipline. In qPCR, DNA amplification is monitored in real time using fluorescent dyes. This improvement provides more accurate and sensitive measurements of e.g. gene expression, pathogen detection, and allelic discrimination, than traditional (end-point) PCR. To achieve high quality results of qPCR, a superb apparatus is needed and this is where you can turn to the Applied Biosystems 7300 Real-Time PCR System.

The 7300 is a very reliable platform with a high quality Peltier-based heating block providing uniform heating (and cooling) of all the wells. It works equally well with both 96-well plates and 0.2 ml tubes, except you must have the converter to utilize both the 96-well format and 0.2 ml tubes. I use the tubes and plates from Applied Biosystems although I have found that Fisher sells the equivalent for cheaper (I haven’t tested them yet). Convenient features include the real-time monitoring of amplification during the run. (Although I am not one to sit and watch the entire run!) I have been using this instrument to detect expression of many protein-encoding genes and to determine drug-induced changes in the expression of these genes.

I typically use Invitrogen products for the isolation of RNA from cultured cells (Trizol) and the Superscript III First-Strand cDNA Synthesis System . I use Applied Biosystems’ Power SYBR® Green 2X Mastermixfor all applications. I have also used Invitrogen’s SYBR Greener mixes and have found similar results as compared to the Power SYBR Mix. I am very curious as to what will happen with the acquisition of Applied Biosystems by Invitrogen and which premix will prevail. The other type of dye chemistry available for use is Applied Biosystem’s TaqMan®, however, since I have never used this chemistry, I cannot offer advice on it.

The 7300 Real-Time software is okay. It takes a little time to learn how to use it but once you are familiar, it is fairly easy to go through and experiments can be set up fairly quickly. Some of the features that I do like are the easy setup of a reaction plate. I’m not fond of the term “detectors” for PCR primers in the software but have adjusted. Also, when you purchase the system you are allowed to install the software on several computers. The downside is that you must input your detectors into each computer that you are working with and permanently save. This can be a problem if multiple labs or multiple users are using the software as there is no way to create subgroups of detectors. Graphs can easily be exported to PowerPoint, however, you cannot change the labeling on the graphs. I have found it easier to just export the data into EXCEL and plot my own graphs.

Overall, it is a powerful system with software bugs that will hopefully be worked out soon.