Polymerase chain reaction cycle

Error Rates in Polymerase Chain Reaction Cycling... 

FIGURE 28.14 The polymerase chain reaction (PCR). Three cycles are shown the target region appears after the third cycle. Additional cycles lead to amplification of the target region. 

FIGURE 13.21 Polymerase chain reaction (PCR). Oligonucleotides complementary to a given DNA sequence prime the synthesis of only that sequence. Heat-stable Taq DNA polymerase survives many cycles of heating. Theoretically, the amount of the specific primed sequence is doubled in each cycle. 

Yoon DS, Lee Y-S, Lee Y, Cho HJ, Sung SW, Oh KW, Cha J, Lim G (2002) Precise temperature control and rapid thermal cycling in a micromachined DNA polymerase chain reaction chip. J Micromech Microeng 12 813-823... 

Figure 40-7. The polymerase chain reaction is used to amplify specific gene sequences. Double-stranded DNA is heated to separate it into individual strands. These bind two distinct primers that are directed at specific sequences on opposite strands and that define the segment to be amplified. DNA polymerase extends the primers in each direction and synthesizes two strands complementary to the original two. This cycle is repeated several times, giving an amplified product of defined length and sequence. Note that the two primers are present in excess.

Polymerase chain reaction (PCR) is one of the most important techniques for rapid bacterial identification. It consists of repeated cycles of enzymatic reactions in a thermal cycler (PCR machine) that copies DNA strands many times. The DNA amplified in one PCR cycle is used as a template for the next cycle. This results in an exponential increase of the desired target... 

The same concept of volumetric in situ heating by microwaves was also exploited by Larhed and coworkers in the context of scaling-up a biochemical process such as the polymerase chain reaction (PCR) [25], In PCR technology, strict control of temperature in the heating cycles is essential in order not to deactivate the enzymes involved. With classical heating of a milliliter-scale sample, the time required for heat transfer through the wall of the reaction tube and to obtain an even temperature in the whole sample is still substantial. In practice, the slow distribution of heat... 

J.R. Uhl, C.A. Bell, L.M. Sloan, M.J. Espy, T.F. Smith, J.E. Rosenblatt and F.R. Cockerill, Application of rapid-cycle real-time polymerase chain reaction for the detection of microbial pathogens the Mayo-Roche rapid anthrax test, Mayo Clin. Proc., 77 (2002) 673-680. 

Figure 13.16 The polymerase chain reaction for the amplification of DNA sequences. DNA is heated to separate the two strands. A primer is attached to the 5 end of each strand and extended using DNA polymerase 1. The two new strands are separated as before and the cycle repeated up to 30 times.

Fig. 9.4 Establishment of the quantitative methylation-specific polymerase chain reaction (MSP) analytical procedure. The ABI PRISM 7900 HT Sequence Detector system was used to perform real-time polymerase chain reaction (PCR) using MSP primers and bisulfite-modified template DNA. Upper panels Setting up the conditions to obtain the standard curves with 50% (A) or 25% (B) sequential dilution of the template. Lower panels The amplification curves on the left represent P-actin, unmethylated, and methylated MSP products, respectively, for reelin (RELN) (C). Amplification curves were compared at the set threshold before 40 cycles. Amplification curves from various samples are shown in the lower panel right (D)...

Fig. 5. Selection scheme for the in vitro selection of RNA libraries. The RNA library is subjected to a selection criterion suitable for the enrichment of functionally active sequences. The few selected individual sequences are amplified by reverse transcription (RT) and polymerase chain reaction (PCR). The PCR-DNA is then subjected to in vitro transcription with T7 RNA polymerase. The resulting enriched and amplified RNA library can be used as the input for the next selection cycle. This process is repeated until active sequences dominate the library. At this point, individual sequences can be obtained by cloning and their sequence can be determined by sequencing...

A novel use of the nuclei isolated from paraffin-embedded tissue is to study a low number of nuclei by polymerase chain reaction (PCR). PCR has the end result of making DNA from a complementary DNA template (2,3). Nuclei can be sorted on the basis of cell cycle, digested to single-stranded DNA, and the desired segment of DNA amplified (4,5). Finally, the isolated nuclei can not only be stained for DNA content, but also for nuclear proteins, proliferation factors, and other nuclear proteins (6-8). 

One method of amplifying DNA in a polymerase chain reaction is through temperature cycling. The double-stranded DNA is denatured between 90°C and 100°C, while the annealing and extension demands lower temperatures. Sufficient amplification of the DNA occurs only after many cycles of temperature change. 

Another complex biochemical reaction is the polymerase chain reaction (PCR). It has also be implemented in liposomes (Oberholzer et al, 1995a). This reachon was interesting from the point of view of vesicle chemistry because the liposomal system had to endure the extreme PCR condihons, with several temperature cycles up to 90 °C (liposomes were practically unchanged at the end of the reaction) and furthermore, nine different chemicals had to be encapsulated in an individual liposome for the reaction to occur. This was carried out by mechanical entrapment from a solution that contained all components only a minimal number of the in situ formed vesicles could entrap all nine components. These odds notwithstanding, there was a significant synthesis. ... 

A thermal cycler (see 8.2.3.2 Polymerase Chain Reaction) is required for the cycle sequencing reaction. For fluorescent cycle sequencing we recommend instruments from Applied Biosystems (e.g. the 16-capillary ABI PRISM 3100 Genetic Analyser) other instruments are available from GC Healthcare and Beckman. For detailed instructions, refer to the respective user s manual or chemistry guide. 

The polymerase chain reaction utilizes a thermostable DNA polymerase to amplify DNA through a series of temperature cycle steps. The key to the specificity of the reaction is the selection of oligonucleotide primers that hybridize to the opposite strands of the DNA being tested, about 400-2000 bp apart. If the sequence of the primers is unique within the genome, and the primers hybridize to the target DNA at a high enough temperature to avoid close matches (various... 

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