Polymerase chain reaction primer extension

Figure 3.25 Polymerase chain reaction. The steps involved in the chain reaction are as follows (i) Incubation of the DNA at a temperature above 90 °C in order to separate the two strands of the DNA duplex, (ii) Cooling of the solution to about 50 °C to allow annealing of the primers to the template (i.e. the nucleotides bind to the template DNA according to the basepairing rules), (iii) Finally, addition of the polymerase and Mg ions to extend the nucleotide primer and complete the synthesis of the complementary DNA, which takes place at about 70 °C. (iv) The sequence (i) to (iii) is repeated to allow another extension to occur many repetitions can be carried out which results in enormous multiplication of the DNA strands. NTPs - deoxyri-bonucleoside triphosphates.

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. 

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... 

Figure 15.5 Polymerase chain reaction step 1, separation of strands by heating (98°C) step 2, anneal primers (60°C) step 3, primer extension by polymerase.

Preparation of Probe. Probes for genomic Southern blots have been made by a variety of methods and must be of high specific activity. A simple method that yields a probe with a very high specific activity utilizes the polymerase chain reaction (PCR).16 An advantage of PCR over methods based on nick translation or primer extension is that only one strand of DNA is synthesized, so that reassociation of denatured single-strand probe in solution is minimized. 

The solid-phase synthesis of DNA oligomers has reached a high level of maturity and is extensively used for the synthesis of primers for polymerase chain reaction or the design of new genes (13). In contrast to the cellular process, the synthetic route forms the polymer in 3 -to-5 direction. The oligomerization is carried out on CPG using a base labile linker. Typically, the nucleobases are introduced as acyl-protected phosphoramidites,... 

Figure 6.8. The First Cycle in the Polymerase Chain Reaction (PCR). A cycle consists of three steps strand separation, hybridization of primers, and extension of primers by DNA synthesis.

PCR (polymerase chain reaction) is a technique in which cycles of denaturation, annealing with primer, and extension with DNA polymerase, are used to amplify the... 

DASH, dynamic allele-specific hybridization AS-PCR, allele-specific polymerase chain reaction AS-PE, allele-specific primer extension APEX, arrayed primer extension FP-TDI, fluorescence polarization template directed dye terminator incorporation MALDI-TOF-MS, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry OLA, oligonucleotide ligation assay RCA, ... 

Figure 5.10 Schematic representation of analysis of single nucleotide polymorphisms (SNPs) by primer extension and MALDI-TOF-MS. The genomic target region is first amplified by the polymerase chain reaction (PCR). An oligonucleotide primer is then annealed immediately adjacent to the polymorphic site. This primer is extended allele-specifically using a DNA polymerase and a nucleotide mix that leads to a termination of the primer extension reaction either after one or two nucleotide additions. The length of the extension products is determined by the allele present in the analyzed sample. In the depicted case, the...

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