The thermal cycler (also known as a thermocycler, PCR machine or DNA amplifier) is a
laboratory
A laboratory (; ; colloquially lab) is a facility that provides controlled conditions in which scientific or technological research, experiments, and measurement may be performed. Laboratory services are provided in a variety of settings: physici ...
apparatus most commonly used to amplify segments of
DNA via the
polymerase chain reaction
The polymerase chain reaction (PCR) is a method widely used to rapidly make millions to billions of copies (complete or partial) of a specific DNA sample, allowing scientists to take a very small sample of DNA and amplify it (or a part of it) ...
(PCR). Thermal cyclers may also be used in laboratories to facilitate other temperature-sensitive reactions, including
restriction enzyme
A restriction enzyme, restriction endonuclease, REase, ENase or'' restrictase '' is an enzyme that cleaves DNA into fragments at or near specific recognition sites within molecules known as restriction sites. Restriction enzymes are one class ...
digestion or rapid diagnostics. The device has a ''thermal block'' with holes where tubes holding the reaction mixtures can be inserted. The cycler then raises and lowers the temperature of the block in discrete, pre-programmed steps.
History
The earliest thermal cyclers were designed for use with the
Klenow fragment of
DNA polymerase I. Since this enzyme is destroyed during each heating step of the amplification process, new enzyme had to be added every cycle. This led to a cumbersome machine based on an automated
pipettor, with open reaction tubes. Later, the PCR process was adapted to the use of
thermostable DNA polymerase from ''
Thermus aquaticus'', which greatly simplified the design of the thermal cycler. While in some old machines the block is submerged in an oil bath to control temperature, in modern PCR machines a
Peltier element
Thermoelectric cooling uses the Peltier effect to create a heat flux at the junction of two different types of materials. A Peltier cooler, heater, or thermoelectric heat pump is a solid-state active heat pump which transfers heat from one si ...
is commonly used. Quality thermal cyclers often contain
silver
Silver is a chemical element with the Symbol (chemistry), symbol Ag (from the Latin ', derived from the Proto-Indo-European wikt:Reconstruction:Proto-Indo-European/h₂erǵ-, ''h₂erǵ'': "shiny" or "white") and atomic number 47. A soft, whi ...
blocks to achieve fast temperature changes and uniform temperature throughout the block. Other cyclers have multiple blocks with high heat capacity, each of which is kept at a constant temperature, and the reaction tubes are moved between them by means of an automated process. Miniaturized thermal cyclers have been created in which the reaction mixture moves via channel through hot and cold zones on a
microfluidic chip. Thermal cyclers designed for
quantitative PCR have optical systems which enable fluorescence to be monitored during reaction cycling.
Modern innovations
Modern thermal cyclers are equipped with a heated lid that presses against the lids of the reaction tubes. This prevents condensation of water from the reaction mixtures on the insides of the lids. Traditionally, a layer of
mineral oil
Mineral oil is any of various colorless, odorless, light mixtures of higher alkanes from a mineral source, particularly a distillate of petroleum, as distinct from usually edible vegetable oils.
The name 'mineral oil' by itself is imprecise, ...
was used for this purpose. Some thermal cyclers are equipped with a fully adjustable heated lid to allow for nonstandard or diverse types of PCR plasticware.
Some thermal cyclers are equipped with multiple blocks allowing several different
PCR PCR or pcr may refer to:
Science
* Phosphocreatine, a phosphorylated creatine molecule
* Principal component regression, a statistical technique
Medicine
* Polymerase chain reaction
** COVID-19 testing, often performed using the polymerase chain r ...
s to be carried out simultaneously. Some models also have a ''gradient'' function to allow for different temperatures in different parts of the block. This is particularly useful when testing suitable annealing temperatures for PCR
primers.
References
External links
OpenPCR an open-source PCR thermal cycler
{{DEFAULTSORT:Thermal Cycler
Molecular biology laboratory equipment
Polymerase chain reaction