Polymerase chain reaction types

The use of agarose as an electrophoretic method is widespread (32—35). An example of its use is in the evaluation and typing of DNA both in forensics (see Forensic chemistry) and to study heritable diseases (36). Agarose electrophoresis is combined with other analytical tools such as Southern blotting, polymerase chain reaction, and fluorescence. The advantages of agarose electrophoresis are that it requires no additives or cross-linkers for polymerization, it is not hazardous, low concentration gels are relatively sturdy, it is inexpensive, and it can be combined with many other analytical methods. 

Electrophysiological studies often demonstrate multiple, asymmetric mononeuropathies, usually axonal in type, that cannot be localized to typical sites of entrapment (Keswani et al. 2002). CSF analysis reveals nonspecific abnormalities, such as elevated protein and mild mononuclear pleocytosis. Polymerase chain reaction (PCR) for CMV DNA and nerve or muscle biopsy may provide more specific diagnostic data (Roullet et al. 1994). 

Pollen analysis in combination with other techniques is still an effective tool for the authentication of the botanical origin of honey (Persano Oddo et ah, 1995 Von der Ohe et ah, 2004). It can distinguish polyfloral and different types of unifloral honeys (Mateo and Bosch-Reig, 1998). It can also indicate the percentages of different nectar contributions in honey products. A polymerase chain reaction technique and an... 

Baba, Y., Tomisaki, R., Sumita, C., Morimoto, I., Sugita, S., Tsuhako, M., Miki, T., and Ogihara, T., Rapid typing of variable number of tandem repeat locus in the human apolipoprotein B gene for DNA diagnosis of heart disease by polymerase chain reaction and capillary electrophoresis, Electrophoresis, 16, 1437, 1995. 

Laboratory confirmation is vital to effective treatment of HSV, especially in individuals in whom a clinical diagnosis cannot be obtained. There are several methods by which a definitive diagnosis may be acquired, and these include virologic typing, serologic diagnosis, rapid point-of-care antigen detection, enzyme-linked immunosorbent assay (ELISA), immunoblot, and DNA polymerase chain reaction.27... 

Molecular methods used to uncover mutations are subject to several variables. The anticoagulants used for blood collection can affect digestion with restriction enzymes and amplification reactions. The type of detergent used in cell lysis can affect amplification of DNA by inhibiting the DNA-amplifying enzyme such as the taq polymerase used in the polymerase chain reaction (116). The control of contamination is crucial in ensuring the quality of results obtained by molecular analysis (117). 

Two principal types of nucleic acid-based methods, nucleic acid hybridization and polymerase chain reaction (PCR), are commonly used for the rapid identification of bacteria. A few other nucleic acid-based methods will also be mentioned. 

Additionally it has been our experience that mass spectrometry as a routine detection/identification technique for bacteria is not well received by microbiologists and clinicians who prefer less expensive, less complicated approaches to bacterial typing and identification, such as methods based on polymerase chain reaction (PCR) and enzyme-linked immunosorbent assays (ELISA). For that reason we have adapted our MS approach to serve as a means of biomarker discovery that feeds candidate proteins or leads into development as PCR targets or other immunoassay techniques. 

CDC Case Definition An illness with acute onset of fever >101°F followed by a rash characterized by firm, deep seated vesicles or pustules in the same stage of development without other apparent cause. Clinically consistent cases are those presentations of smallpox that do not meet this classical clinical case definition (1) hemorrhagic type, (2) flat type, and (3) variola sine eruptione. Laboratory criteria for diagnosis is (1) polymerase chain reaction (PCR) identification of variola DNA in a clinical specimen, or (2) isolation of smallpox (variola) virus from a clinical specimen (Level D laboratory only confirmed by variola PCR). 

Cytokine profiling has also been measured as a function of changes in cytokine mRNA expression using either reverse transcription polymerase chain reaction (RT-PCR) [87, 91-93] or ribonuclease protection assay (RPA) [94-97], Measurement of cytokine transcripts by RT-PCR revealed that prolonged exposure to TMA induced increased levels of IL-4 mRNA expression compared with treatment with DNCB [87,92-93]. However, expression of the type 1 cytokine IFN-y by DNCB-activated LNC was variable and failed to discriminate between contact and respiratory allergens [87,91,93). A similar profile was observed for freshly isolated tissue analyzed by RPA. This somewhat less... 

Phosphoinositase C (i.e. phosphoinositide-specific phospholipase C [PLC]) enzymes are found in the vast majority of mammalian cells. Molecular cloning of these enzymes, analysis of their predicted amino acid sequences and immunological cross-reactivity indicate that at least three major forms of the enzyme exist PLC-/I, -8 and -y. Each of these enzyme types is encoded by a distinct gene. More recent experiments using the polymerase chain reaction and molecular cloning have revealed even greater enzyme di-... 

B3. Begovich, A. B., and Erlich, H. A., HLA typing for bone marrow transplantation. New polymerase chain reaction-based methods. JAMA, J. Am. Med. Assoc. 586-591 (1995). 

List of Abbreviations PCR, polymerase chain reaction RT-PCR, reverse transcription polymerase chain reaction DNA, deoxyribonucleic acid RNA, ribonucleic acid RNase, ribonuclease mRNA, messenger RNA GABAa, y-aminobutyric acid type A cRNA, copy RNA dNTPs, deoxy nucleoside triphosphates MMLV, Mouse Moloney murine leukemia vims RT, reverse transcriptase bp, base pair Tm, melting temperature DEPC, diethylpyrocarbonate OD, optical density mL, milliliter SA-PMPs, streptavidin paramagnetic particles dT, deoxy thymidine DTT, dithiothreitol DNase, deoxyribonuclease RNasin, ribonuclease inhibitor UV, ultraviolet TBE, Tris-borate, 1 mM EDTA EDTA, ethylenediaminetetraacetic acid Buffer RET, guanidium thiocyanate lysis buffer PBS, phosphate buffered saline NT2, Ntera 2 neural progenitor cells... 

List of Abbreviations GABA, gamma-aminobutyric acid type A 5HT3, 5-hydroxytryptamine type 3 SDM, site-directed mutagenesis PCR, polymerase chain reaction TRCP, targeted random chimera production SDS, sodium dodecyl sulphate... 

Nevas, M., Hielm, S., Lindstrom, M., Horn, H., Koivulehto, K. and Korkeala, H., High prevalence of Clostridium botulinum types A and B in honey samples detected by polymerase chain reaction, Int. J. Food Microbiol., 72, 45-52, 2002. 

Two types of screening procedures are used to identify ES cell clones carrying a targeted integration of the construct DNA polymerase chain reaction (PCR) and Southern blot analysis (Southern, 1975). Both methods rely upon the specific juxtaposition of vector components and target locus sequences after homologous recombination. 

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