Thermal cycler

Template:Short descriptionThe thermal cycler (also known as a thermocycler, PCR machine or DNA amplifier) is a laboratory apparatus most commonly used to amplify segments of DNA via the polymerase chain reaction (PCR).<ref>Template:Cite journal</ref> Thermal cyclers may also be used in laboratories to facilitate other temperature-sensitive reactions, including restriction enzyme digestion or rapid diagnostics.<ref>Template:Cite journal</ref> 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

File:Baby Blue - a prototype polymerase chain reaction (PCR), c 1986. (9663810586).jpg

Baby Blue, a prototype automated thermal cycler built around 1986

File:Primitive PCR machine for scrap.JPG

A very early PCR machine which, rather than cycling through different temperatures, uses three different water baths at constant temperatures between which samples are moved with a robotic arm

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 is commonly used. Quality thermal cyclers often contain silver 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 innovation

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 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.<ref>Template:Cite news</ref>

Some thermal cyclers are equipped with multiple blocks allowing several different PCRs 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.

File:Working model of thermal cycler machines v2.webm

CitationClass=web }}</ref> uses this technology.

	Peltier based	Resistive Heating Element	Air Chamber
Commercially available machines	citation	CitationClass=web }}</ref>	citation	CitationClass=web }}</ref>	citation	CitationClass=web }}</ref>
Thermal Elements	Peltier Element	Resistive Heating	Resistive Heating
Lid Heater	yes	no	no
Heating Rate	<10 °C/Sec	<4 °C/Sec	>15 °C/Sec
Cooling Rate	<5 °C/Sec	<2.5 °C/Sec	>20 °C/Sec
Gradient Temperature	yes	no	no
Temperature uniformity	0.15 °C	0.5 °C	0.02 °C
Power consumption	<850 W	<72 W	<520 W

External links

• OpenPCR, an open-source PCR thermal cycler