Molecular biology techniques polymerases

The EMBL vector systems have been subjected to many modifications to enhance their usefulness. The most useful modifications involve the addition of more restriction enzyme recognition sites flanking the stuffer fragment and the addition of T3 and T7 RNA polymerase promoters. The T3 and T7 RNA polymerase promoters are used to synthesize 32P-labeled RNAs that can be used as probes for the identification of overlapping A clones. Many of the A vectors described above are available from or have been developed by commercial vendors. As in the case with many basic molecular biology techniques, the components needed to construct a specific recombinant phage library (A vector arms, in vitro packaging extracts, etc.) and even the library can be obtained from commercial sources. 

Molecular biology techniques, such as Southern blot analysis and polymerase chain reaction (PCR), have been used in the diagnosis of hemoglobinopathies (see the following discussion). 

Methods for the detection of specific nucleic acid sequences are much more sensitive with respect to the detection of proteins, since polymerase chain reaction (PCR) represents a very powerful amplification tool for nucleic acids. The fact that aptamers are made of DNA or RNA offers the opportunity to combine the detection of proteins with the amplification potentialities that until some years ago were restricted to the detection of nucleic acids. PCR has been used alone or combined with other molecular biology techniques for nucleic acid manipulation (proximity ligation or exonuclease protection) to develop extremely sensitive assays for the detection of a variety of proteins. 

PCR Lab-on-Chip devices are miniaturized fluidic systems for fast DNA amplification. The polymerase chain reaction (PCR) is a molecular biological technique for addressing and exponentially amplifying a DNA fragment. The PCR process is based on tenperature cycles with an enzymatic amplification step. It works without the use of a living cell. For PCR, a liquid mixture of an analyte... 

Progress in molecular biology has provided a new perspective. Techniques such as the polymerase chain reaction and single-strand conformation polymorphism analysis have greatly facilitated the molecular analysis of erythroenzymopathies. These studies have clarified the correlation between the functional and structural abnormalities of the variant enzymes. In general, the mutations that induce an alteration of substrate binding site and/or enzyme instability might result in markedly altered enzyme properties and severe clinical symptoms. 

The most widely used amplification technique is the polymerase chain reaction. Hardly a research study in molecular biology involving DNA amplification has been performed without using PCR. Many modifications of PCR have been described. Before we review these modified techniques, it would be appropriate to briefly review the PCR methodology. 

A technique that has revolutionized the field of molecular biology during the last decade is the polymerase chain reaction (PCR). This method, conceived by Kary B. Mullis (1944—) in 1983, involves making multiple copies of fragments of DNA in a short time. Mullis received the 1993 Nobel Prize in chemistry for his work. PCR mimics DNA s natural ability to replicate itself It involves three basic steps conducted at different temperatures. In the first step, a mixture of DNA and other basic PCR ingredients is heated in a test tube to approximately 90°C. At this temperature, the DNA strands unwind. The second step involves lowering the temperature to around 55°C, which allows special enzymes called... 

Kaiy Mullis and others at Cetus Corporation in Berkeley, California, invented a technique for multiplying DNA sequences in vitro by, the polymerase chain reaction (PCR). PCR has been called the most revolutionary new technique in molecular biology in the 1980s. Cetus patented the process, and in the summer of 1991 sold the patent to Hoffmann La Roche. Inc. for 300 million. 

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