The thermocyclor (also called thermal cyclor or machine PCR) is an apparatus automating the reaction of PCR " classique" or " not final". The apparatus is provided with a thermal block (" thermal block") with holes where one can insert the tubes containing the reactional mixture of the PCR.
Certains model makes it possible to carry out heat gradients, on a side of the thermal block to the other. This function is used when one wants to find the best temperature of hybridization for a couple of starters given.
The modern aircraft are equipped with a heating roof , which presses on the caps of the tubes and avoids the evaporation of the reactional mixture and avoids having to put a mineral oil film at the top of the reactional medium in the tube.
A keyboard composed of some keys and a screen with liquid crystals make it possible to enter of the Programme S the mêmoire of the machine.
The first price for a thermocyclor is approximately 2.500 US$.
Program PCR
Program example:- 95°C during 10 to 15 minutes (in the event of enzyme " Hot Start ")
- approximately 40 cycles made up of:
- 95°C during 30 seconds (stage of denaturation)
- Tm +2 °C (by ex: 55°C) during 1 minute (stage of Hybridization or " annealing")
- 72°C during 1 minute (stage of elongation)
- 72°C during 5 minutes
- 40°C during 1 minute (" cooling")
Note: the stage of elongation is function length awaited of the fragment to be amplified. It is considered that the Taq polymerase synthesizes approximately 1000 bases/minute. For an awaited length of 500 Pb, 30 seconds of elongation are sufficient.
Program sequencing
Generally only one score of cycles includes/understands.
Program denaturation
The program generally called " DENAT". It consists of a single stage with 95°C during 10 or 15 minutes. For the end of the cycle, it is necessary to be present to quickly leave the bars samples to quickly plunge them in the ice. Attention, the block of the thermocyclor is hot, one should not be burned.
Crocodile
In the years 1980, the crocodile (still called croco affectionately) was one of the first apparatuses to being sold at affordable prices and having known of this fact a broad diffusion. It used tubes of 0.5 ml. It did not have a heating roof. It was necessary to put a mineral oil film to avoid the evaporation of the reactional medium. It did not use the Peltier effect for cooling, but a ventilator.
Minicycler
The MiniCycler is produced by MJ Research. It has 16 well and uses the Peltier effect to cool the tubes.
PTC
The series of the PTC is produced by MJ-Research. They are thermocycleurs with Peltier effect with a heating roof. They are provided with a block for 96 tubes of 0,2 ml. The variations in temperature reach 3°C a second.- the PTC 100
- the PTC 200
Mastercycler 5330
It is a thermocyclor produced by Eppendorf.
PCR in real-time
See Apparatus of PCR in real-time
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