Polymerase chain reaction PCR amplification

HCV and HIV-1). The bDNA assay53 is being much employed for the quantification of messenger RNA. Moreover, for the detection of viral and pathogenic disorders based on alkaline-phosphatase-sensitive dioxetanes, several assay methods are available these include the Polymerase-Chain-Reaction (PCR) amplification, probe ligation, strand-displacement amplification and the ligase chain reaction53. 

Although a number of mammalian retrotransposons have been discovered by chance, a variety of approaches have been developed specifically to survey the genome for the presence of retrotransposons as well as to facilitate their isolation and characterization. Of the three methods outlined here in detail, two (conserved restriction sites and phylogenetic screening) could lead to isolation of all three types of retrotransposons, whereas the third method [polymerase chain reaction (PCR) amplification of reverse transcriptase] will exclude the SINEs. An unrelated method that has been particularly useful for isolation of SINEs7 will not be detailed. 

The authors describe an ultrasensitive method that measures RT activity. The assay adopts polymerase chain reaction (PCR) amplification for detecting the cDNA product of the reaction, and therefore was named Amp-RT (3,4). Amp-RT measures the ability of a sample to produce a DNA copy of a known heteropolymeric RNA template by extending a complementary DNA oligoprimer. The RNA template used in Amp-RT is a sequence from the genome of the encephalomyocarditis virus. This chapter describes in detail the Amp-RT method and its use as (1) a qualitative assay for the generic detection of retroviruses, (2) a quantitative method to measure virus loads of the human immunodeficiency virus type 1 (HIV-1), and (3), a screening method for susceptibility of HIV-1 to RT inhibitors. 

The TRAP (telomeric repeat amplification protocol) assay is a widely used method for detection of telomerase activity. This technique measures the telomerase activity present in cell extracts. Briefly, cellular extract containing telomerase activity is incubated with a telomeric substrate (a short strand of DNA onto which the telomerase wiU. attach the telomeric repeats) followed by polymerase chain reaction (PCR) amplification of the elongated telomere. Detection of the PCR product is by a number of methods, including gel electrophoresis, radiometric detection, ELISA, and real-time PCR detection. ... 

In addition to functional assays for CYP2D6, polymerase chain reaction (PCR) amplification assays have been developed for the detection of specific mutations in the CYP2D6 gene (table 1). Additional genotyping... 

The dot-blot shown below examines DNA from a child with ambiguous genitalia after polymerase chain reaction (PCR) amplification and hybridization with DNA probes from the X and Y chromosome. In this case, the Y chromosome probe is from the SRY region of Yp that has recently been characterized as the male-determining region. DNA from control male and female patients is also applied to the dot-blot. Based on the dot-blot results, which is the most likely conclusion ... 

Takahashi Y, Itami T, Maeda M, et al. Polymerase chain reaction (PCR) amplification of bacilliform virus (RV-PJ) DNAin Penaeus japonicus Bate and systemic ectodermal and mesodermal Baculovirus (SEMBV) DNA in Penaeus monodon Fabricius. J Fish Diseases 1996 19 399-403. 

HPLC life science applications focus on the separation, quantitation, and purification of biomolecules such as proteins, peptides, amino acids, nucleic acids, nucleotides, and polymerase chain reaction (PCR) amplification products.31 34 These are diversified and active research areas in medical research and drug discovery. 

Several examples of work in this field have been reported for multiple-sample polymerase chain reaction (PCR) amplifications (464 166) and electrophoretic analysis in a microchip. In one example, the PCR products from four DNA samples were analyzed by microchip gel electrophoresis (466). The ability to analyze a large number of DNA samples in this format was presented. Run-to-run reproducibility in the sizing microchip was very good percent relative standard deviation n = 8 for migration times was better than 0.3%. The accuracy of the 500-base-pair sizing was greater than 98% (466). 

To this end, cDNA libraries, prepared from cells or tissues known to be rich in certain receptors, were screened by low stringency hybridization, or were used for polymerase chain reaction (PCR) amplification of candidate genes using degenerate primers. Proof of function was obtained after the... 

To establish whether differences in toxin profiles are due to genetic variability between strains of the same species, DNA analyses are required. Nevertheless, polymerase chain reaction (PCR) amplification of ribosomal DNA subunits and internal transcribed spacers of Dinophysis spp. have shown very little variability in this part of the genome commonly used to separate species and strains of other microalgal species [139-142] sequences of plastid DNA show no differences among the phototrophic Dinophysis spp. tested, or between these and the plastid sequences of some small (8-15 am) cryptophytes of the genus Teleaulax [74]. As a result, DNA probes able to separate Dinophysis at the species level and to And differences between strains of the same species are still on a developmental stage. 

SEB. Therefore, serum antibody titers are of little diagnostic value. If actual bacterial involvement is suspected, and if cultures can be obtained, the detection of extremely minute quantities of potentially toxigenic strains is possible by using (1) polymerase chain reaction (PCR) amplification and (2) toxin gene-specific oligonucleotide primers. The results from both methods are rapid, allowing quantitative or qualitative measurements in less than 24 hours. 

The process is initiated with a random library of linear oligonucleotides (usually, 10 to 10 ) consisting of linear nucleic acids comprising a random sequence embraced by a 5 and a 3 nucleic acid sequence of defined composition. An RNA-searched aptamer involves the primary transcription of the DNA library into an RNA pool followed by passing the library through a separating matrix that includes the target substrate. The few nucleic acids that reveal affinity toward the substrate (or some nonspecific nucleic acid adsorbents) bind to the separation matrix, while most of the library components are washed off. The elution of surface-bound nucleic acids followed by their polymerase chain reaction (PCR) amplification yields a mixture of nucleic acids... 

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