Apparatus of PCR in real-time
systems with capillary tubesThe company Roche markets Light-cyclerTM. Up to 32 capillary tubes, out of glass, of a capacity of 20 µl, is related to a " carousel" , turning in an obscure room of circular form. The stages of temperature are carried out by the means of a current of hot air. The apparatus is able to raise or lower its temperature of room of 20°C a second.
Il has there a lamp of excitation diffusing a light of 530 Nm wavelength and an integrator able to recover the signal corresponding in the light of emission of fluorescence generated to 5 different wavelengths.
La software continuation makes it possible to make quantification absolutely or in a relative way.
Les the most current apparatuses is the versions light to cycle 2.0.
Systems with plates 96 wellsThe systems of PCR real-time by plate 96 wells are most widespread within the community of user. The consumable ones are available at much supplier, their use is simple as well as the handling of the plates. The sticking plastics and films are of optical quality not to disturb detection by parasitic phenomena. The recent machines progressed in boarding and the descent times in temperature of the thermocycles, thus using news more tolerant Taq polymerase towards these fast changes of temperature. In the same way, the last machines propose more channels of detection to leave the possibility to the user of carrying out broader crossings of Fluorochromes (between 4 and 5 different possible colors per machine).
Ci below an not-exhaustive list of various manufacturers and their respective machine
Applied Biosystems : the ABI PRISM 7700,7900 and 7000
- BioRad with iCycler IQ, the IQ5 (5 colors), Mini Opticon, Chromo4 (ex-MJ research)
- Cepheid: SmartCycler (16x6 tubes)
- Eppendorf: Realplex (2 or 4 colors)
- Rock: LightCycler 480
- Stratagene: the MX-4000 and MX-3000 and 3005
System with tubes PCRCorbett Research markets Rotor-Gene 3000 and 6000, of the themocyclers with a rotor which turns in order to read the various wells. They can read 36 or 72 tubes, according to the rotor. Like LightCycler, it functions with pulsated air. It is an apparatus much simpler (and thus more reliable) than those with plates 96 wells because they use a system of sets of mirrors which move in order to read fluorescence. Rotor-Gene 6000 received the price Frost & Sullivan 2006 (Technology Innovation Award, North America)
Developed by the company GeneSystems (Bruz, France) which markets an automat of qPCR (GeneDiscCycler) functioning with a disposable disc ready with employment (GeneDisc). Each GeneDisc contains 36 wells pre-filled by the industrialist with the reagents necessary according to the type of amplified DNA. These 36 wells form 6 identical sectors allowing an analysis by sector. The extract of DNA to be analyzed is directly charged in a sector and it is distributed in the 6 wells (2 positive witnesses, 3 repetitions of the analysis and 1 negative witness). This automat is coupled with another automat of extraction of DNA, GeneExtract.
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