Polymerase chain reaction thermocyclers

Polymerase chain reaction (PCR) The process by which a specific sequence of DNA can be amplified (copied many times) in vitro. It requires a pair of primers and template DNA, thermostable DNA polymerase (e.g. Taq polymerase), deoxynucleotide triphosphates and a thermocycler. The process can amplify large... 

Oda RP, Strausbauch MA, Huhmer AF, Borson N, Jurrens SR, Craighead J, et al. Infrared-mediated thermocycling for ultrafast polymerase chain reaction amplification of DNA. Anal Chem 1998 70 4361-4368. 

Huhmer AF, Landers JP. Noncontact infrared-mediated thermocycling for effective polymerase chain reaction amplification of DNA in nanoliter volumes. Anal Chem 2000 72 5507-5512. 

Figure 5-17. Typical temperature profile of a polymerase chain reaction. The polymerase chain reaction is carried out in a thermocycler where the temperature is controlled by a microprocessor unit. The temperature regime is varied so that the double stranded DNA is...

Small dimensions are also advantageous with respect to heat transfer in reactions such as the polymerase chain reaction (PGR) that are highly dependent on rapid temperatme changes and that are frequently used as sample preparation step prior to biosensor analysis. Reactions taking tjq)ically in the order of hours in a standard thermocycler have been reduced to a few minutes in microchannels [82]. 

Thermocycler. Any themocycler can be used for Polymerase Chain Reaction (PCR). We regularly use the Smart-cycler real-time PCR machine (Cepheid), which allows us to monitor the progress of PCR amplification and decide when to stop the reaction. 

Polymerase chain reaction (PCR) is a powerful method for DNA amplification (i), which is routinely used for detection and analysis of DNA. Many commercial thermocyclers employ a Peltier chip (2-4), which is a solid-state technology that can be used for heating or cooling. Peltier systems are widespread and provide a... 

Amplify the sequence of the gene of interest by polymerase chain reaction (PCR) with the Pwo DNA polymerase using specific primers containing a restriction site for BssHII on 5 end of the sense primer and a Sail restriction site on the 2> end of the antisense primer. Mix 10 ng of template DNA with the specific primers to obtain a final concentration of 800 nM in a volume of 25 pL. Add 25 pL of Pwo Master Mix and incubate on a thermocycler for 30 cycles of denaturation/anneahng/ elongation following the manufacturer s instructions. 

Figure 4 Polymerase chain reaction (PCR). (a) Principle of DNA amplification in PCR (b) a typical thermocycle in PCR. (Adapted from R.A. Gibbs. Anal. Chem. 62 1202-1214,1990. With permission.)...

Polymerase chain reaction (PCR) consists of repeated cycles of in vitro DNA synthesis from a template by using two prim oligonucleotides that hybridize to opposite strands of the ftragment to be amplified. The use of thermostable Taq DNA polymerase enables to run the required steps of denaturation, annealing and DNA chain elongation in an automatic thermocycler, now commercially available from several sources, and thus greatly reduce the time and effort. Recent modifications of the technique allows amplification of DNA fragments with the use of only one specific primer. The other primer is selected to be complementary to sequences in... 

SchneegaB I, Kohler JM (2001) Flow-through polymerase chain reactions in chip thermocyclers. Rev Mol Biotechnol 82 101-121... 

The polymerase chain reaction (PCR) is a means to amplify a given stretch of nucleotides to determine their relative concentration in different cell samples, or to produce usable quantities of a sequence of nucleotides for DNA cloning. The technique allows for the amplification of a few copies (in theory, only one copy is needed) of a specific piece of DNA into perhaps billions of copies in a relatively short time. The procedure is relatively inexpensive, requiring only a thermocycler and the appropriate enzymes and reagents for processing. The first description of PCR was published in 1985 (Saiki et al., 1985), and has proven to be such an important technique that the Nobel Prize in 1993 was awarded to Kary Banks MuUis for its discovery. 

The accuracy of Watson and Crick s description of how the DNA double helix unfolds to rephcate and self-assemble afterwards provided another exciting tool for bioinspired materials, namely a way of controlling self-assemblage. The possibility of controlhng the assembling and disassembling cycles of DNA molecules became a reality with the introduction of the polymerase chain reaction (PCR) as a way to synthesize novel DNA molecules. With a very small amount of DNA, we are now able to replicate it billions of times in a machine called a thermocycler and use the synthetic DNA to transform living cells or to produce completely different materials. Kary Mulhs won the Nobel Prize of Chemistry in 1993 for the invention of PCR. 

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