Polymerase chain reaction selection

Mueller, C.G., Rissoan, M.C., Salinas, B., Ait-Yahia, S., Ravel, O., Bridon, J.M., Briere, F., Lebecque, S., and Liu, Y.J. (1997). Polymerase chain reaction selects a novel disintegrin proteinase from CD40-activated germinal center dendritic cells. J. Exp. Med. 786 655-663. 

Cuypers, H. T. M etal. (1992). Storage conditions of blood samples and primer selection affect the yield of cDNA polymerase chain reaction products of hepatitis C virus. J. Clin. Microbiol 30, 3220-3224. 

It s also possible to select your DNA before you put it in the vector. If you know the sequence (or even part of it), DNA pieces (from genomic DNA or cDNA) with this sequence can be purified on a gel and identified by hybridization to an oligonucleotide using a Southern blot. Alternatively, if you know the sequence of the ends of your DNA, you can amplify it specifically by the polymerase chain reaction. There are lots of clever ways to find your DNA. 

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 set of sense and antisense primers should be selected to synthesize specific cDNAs and also to detect the amplified messages of the genes, complementary to their specific gene sequences. It is important to consider the following points while designing the primers for reverse transcriptase and polymerase chain reaction ... 

PTT is based on a combination of reverse-transcription polymerase chain reaction (RT-PCR) and linked in vitro transcription and translation (Fig. 3C). This combination of procedures can selectively detect translation-terminating or nonsense mutations. Unfortunately, it does not find missense mutations, which may be etiologic, depending upon location. 

Advances in biotechnology have been tremendous. There are many issues in biochemistry and biotechnology that have not yet been completely elucidated however, progress has been made. For example, antibiotics have evolved, the polymerase chain reaction has been developed, and DNA sequencing, genomics, proteomics, combinatorial synthesis, and selective complexation and recognition chemistry have all advanced. 

Examination of stained specimens by microscopy or simple examination of an uncentrifuged sample of urine for white blood cells and bacteria may provide important etiologic clues in a very short time. Cultures of selected anatomic sites (blood, sputum, urine, cerebrospinal fluid, and stool) and nonculture methods (antigen testing, polymerase chain reaction, and serology) may also confirm specific etiologic agents. 

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