Polymerase chain reaction applications, generally

Apart from the traditional organic and combinatorial/high-throughput synthesis protocols covered in this book, more recent applications of microwave chemistry include biochemical processes such as high-speed polymerase chain reaction (PCR) [2], rapid enzyme-mediated protein mapping [3], and general enzyme-mediated organic transformations (biocatalysis) [4], Furthermore, microwaves have been used in conjunction with electrochemical [5] and photochemical processes [6], and are also heavily employed in polymer chemistry [7] and material science applications [8], such as in the fabrication and modification of carbon nanotubes or nanowires [9]. 

The polymerase chain reaction (PCR), developed by Mullis, is a simple and most effective way of amplifying, i.e. producing multiple copies of, a DNA sequence. It finds applications in all sorts of areas not immediately associated with nucleic acid biochemistry, e.g. genetic screening, medical diagnostics, forensic science, and evolutionary biology. The general public is now well aware of the importance... 

Zoh GJ, Melchers WJ, Kopecka H, Jambroes G, van der Poel HJ, Galaraa JM. General primer-mediated polymerase chain reaction for detection of enteroviruses application for diagnostic routine and persistent infections. J Clm Microbiol 1999 30 60-5. 

In this chapter we describe methods used to analyze biological samples by sequencing analysis. The main area focuses on methods used typically in routine laboratories. The applications described are examples that represent the wide field of applications for sequencing in daily analysis work. All methods described here are based on polymerase chain reaction (PCR), which is described in Chapter 2. Generally, the success of analysis depends on the correct sampling and storage, the DNA content of the sample, and the correct DNA extraction method. 

The utilization of classical polystyrene particles or hydrophobic latexes for protein concentrations can induce undesirable phenomena such as protein denaturation and low concentration yields, on account of the high adsorption affinity between both species, which may lead to low desorbed amount. In addition, the use of such hydrophobic colloids in the polymerase chain reaction (PCR) nucleic-acid amplification step generally leads to total inhibition of the enzymatic reaction. The inhibition phenomena can be attributed to the denaturation of enzymes adsorbed in large numbers onto hydrophobic colloids. The utilization of hydrophilic and highly hydrated latex particles (irrespective of temperature) is the key to solving this problem by suppressing the inhibition of enzyme activity. The purpose of this stage is then to focus on the potential application of thermally responsive poly(NIPAM) particles for both protein and nucleic acid concentrations. 

The key to this work is DNA—the chemical fingerprint present in every tissue of every individual. Although the general structure of DNA is the same from one person to another, evidence for familial ties is present in the detailed sequence of each person s DNA. With the use of relatively simple chemistry—involving fluorescent dyes or radioactive isotopes, enzymes, gel electrophoresis, and a process called the polymerase chain reaction (PCR) that earned its inventor the 1993 Nobel Prize in Chemistry (Section 25.8)—it is now easy to synthesize millions of copies from a sample of DNA and to sequence it rapidly and conveniently. Application of these tools to comparison of DNA samples from victims and relatives provides hope that, at least in some cases, the gap between family members will be closed. 

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