Bio-Rad Real Time PCR Systems are designed for optimal performance, reliability and ease of use. These systems are available in a variety of formats to meet the needs of all your molecular biology applications. It use the same revolutionary technology that made Bio-Rad’s thermal cycler the industry standard. They combine outstanding thermal stability with high sensitivity and accuracy. This kit contains all the necessary reagents and consumables to perform a real-time PCR assay.
The BioRad Real-Time PCR system uses SYBR Green I dye, which binds to double-stranded DNA when bound by the polymerase chain reaction (PCR) enzyme, TaqMan® probes. The dye fluoresces when excited by specific light, generating an emission spectrum that can be distinguished from background emission. The fluorescent signal is measured using an instrument containing a laser source and PMT detectors. The intensity of the fluorescent signal is proportional to the amount of PCR product amplified in each reaction.
Real-time PCR is a high-throughput, quantitative assay for detecting and quantifying specific DNA or RNA sequences. Real-time PCR is performed in a closed reaction vessel (real-time cycler) with fluorescence detection by the fluorophore incorporated into the primer. The amplification products are detected as they accumulate during the PCR, so that data can be obtained in less than one hour. It is used to detect small numbers of target sequences (DNA or cDNA), including those that are present at low frequency in a complex background of genomic DNA. Real-time PCR has been used for a wide variety of applications, including genotyping, mutation screening, microbial identification and quantification, gene expression studies and drug resistance detection.
Bio-Rad recommends using Perkin Elmer GeneAmp 5700 Sequence Detection System for real time PCR. The equipment includes a thermal cycler, software and a dedicated lysing station. The system can be used with 96-well or 384-well plates.
The following components are required:
- Real time thermocycler (Perkin Elmer ABI Prism 7700 Sequence Detection System)
- Microscope (Leica DMIRB microscope with AxioCam MRc5 camera)
- Injector (Hamilton syringe or Eppendorf Nanoject)
- Plate washer (Eppendorf Multidrop Combi).
The first thing you need to consider is the safety and security of your PCR machine. If it isn’t safe and secure, then it is not worth buying at all. The safest PCR machine would be one that has been approved by the FDA, or any other government regulatory agency such as OSHA. This means that it has passed certain safety tests and has been deemed safe to use in a laboratory setting. You also want to make sure that your PCR machine comes with an interlock system which will prevent anyone from entering the room while it is on. There are many types of interlock systems available, but they all serve the same purpose: keeping people out of danger while a hazardous chemical is being used in the lab.
Another thing that you need to consider when buying a PCR machine is how large your lab is going to be. Most labs only have one person working in them at any given time, although some may have multiple people depending on how big their budget.