Polymerase chain reaction denaturation

Imai M, Hoshi T, Ogawa K. K-ras codon 12 mutations in biliary tract trrmors detected by polymerase chain reaction denaturing gradient gel electrophoresis. Cancer. 1994 73 2727-2733. 

Hutzler, M., Geiger, E., Jacob, F. (2010). Use of PCR-DHPLC (Polymerase chain reaction-denaturing high performance liquid chromatography) for the rapid differentiation of industrial Saccharomyces pastorianus and Saccharomyces cerevisiae strains. Journal of the Institute of Brewing, 116, 464-474. 

G. Muyzer, E. C. de Waal, and A. G. Uitterlinden, Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for I6S rRNA. Appl. Environ. Microbiol. 59 695 (1993). 

Stothard, J.R., Frame, IA. and Miles, MA. (1997) Use of polymerase chain reaction-based single strand conformational polymorphism and denaturing gradient gel electrophoresis methods for detection of sequence variation of ribosomal DNA of Trypanosoma cruzi. International Journal for Parasitology 27, 339-343. 

Such renaturation or annealing of complementary DNA strands is an important step in probing a Southern blot and in performing the polymerase chain reaction (reviewed in Chapter 7). In these techniques, a well-characterized probe DNA is added to a mixture of target DNA molecules. The mixed sample is denatured and then renatured, When probe DNA binds to target DNA sequences of sufScient complementarity, the process is called hybridization. 

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. 

Steps in the polymerase chain reaction (PCR). The DNA to be amplified is denatured and annealed with two oligonucleotides that flank the region of interest. These oligonucleotides (or primers) are extended. Extension continues to the ends of the DNA strands. The products are again denatured and annealed to primers for a second round of extension. This process of denaturation, annealing, and primer extension is repeated many times. The primary product of the reaction is duplex DNA, bounded by the sequences of the primers. (From J. L. 

Figure 4.2 Polymerase chain reaction (PCR) (Lottspeich, 1998). The PCR cycles between a denaturing step to obtain single-stranded DNA, an annealing step for the primer attachment to the template DNA, and a polymerization step, in which the heat-stable polymerase elongates the corresponding strand using the primers as starting points.

Immunocapture-polymerase chain reaction (IC-PCR) is a synthesis of two commonly used diagnostic tools. This method exploits the high-affinity binding of antibodies to provide a facile method of purification, usually from a complex matrix, supplying the substrate for PCR detection. PCR exponentially amplifies a deoxyribonucleic acid (DNA) template in a temperature-dependent fashion by the annealing of oligonucleotide primers, enzymatic extension of bound primers by a heat-stable polymerase, followed by denaturation of... 

ECL has also been applied to the analysis of polymerase chain reaction (PCR) products [53,55], PCR is first used to amplify the specific genes by use of two primers, one of which is biotinylated. The double-stranded DNA is then captured on streptavidin-coated magnetic beads and washed with an alkaline solution to denature and separate the strands. The particle-bound, single-strand DNA is used to capture products hybridized with Ru-labeled complementary... 

Polymerase chain reaction (PCR) offers a convenient method of amplifying the amount of a DNA segment (Mullis et al, 1994 Newton and Graham, 1997 Saiki et al, 1988 Timmer and Villalobos, 1993). In this technique, the DNA synthesizing preparation contains denatured DNA with the segment of interest that serves as template for DNA polymerase (DNA-directed DNA polymerase), two oligonucleot-... 

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