Polymerase chain reaction variations

Stothard, J.R., Frame, IA. and Miles, MA. (1997) Use of polymerase chain reaction-based single strand conformational polymorphism and denaturing gradient gel electrophoresis methods for detection of sequence variation of ribosomal DNA of Trypanosoma cruzi. International Journal for Parasitology 27, 339-343. 

A variation on the theme of conventional assay uses both lanthanide-labeled and biotin-labeled single strands to form split probes for sequence of target strands (Figure 12).120 When both of these bind to DNA, the complex binds (via the biotin residue) to a surface functionalized with streptavidin, immobilizing the europium and allowing assay to be carried out. This approach is already very sensitive to DNA sequence, since both sequences must match to permit immobilization of the lanthanide, but can be made even more sensitive by using PCR (the polymerase chain reaction) to enhance the concentration of DNA strands. In this way, initial concentrations corresponding to as few as four million molecules can be detected. This compares very favorably with radioimmunoassay detection limits. 

Heniford, B W., Shum-Siu, A, Leonberger, M., and Hendler, F J (1993) Variation m cellular EGF receptor mRNA expression demonstrated by in situ reverse transcriptase polymerase chain reaction. Nucleic Acids Res 21,3159-3166... 

Analysis of DNA Sequence Variation at Population Level by Polymerase Chain Reaction and Denaturing Gradient Gel Electrophoresis... 

Meyer W Mitchell TG (1995) Polymerase chain reaction fingerprinting in fungi using single primers specific to minisatellites and simple repetitive DNA sequences strain variation in Cryptococcum neoformans. Electrophoresis 16 1648-1656. 

Molecular techniques allow the evaluation of molecular-level (DNA and RNA) variation in populations. Most techniques use the polymerase chain reaction (PCR) to amplify short nucleotide sequences from small amounts of tissues. With DNA techniques, one can examine neutral markers (non-coding regions widely dispersed over the entire genome) or patterns at coding loci (regions that are essential for functional differences in important proteins or for gene expression). Also, mitochondrial DNA (mtDNA) (see Lecrenier and Foury, 2000) has... 

The value of the RFLP technique lies in its ability to demonstrate allelic variation within specificities that do not show variation at the serological level (K13). While the RFLP technique is valuable in determining allelic variation, it is subject to certain limitations. The technique is labor intensive and time consuming, and does not lend itself to routine application. Moreover, the procedure requires large amounts of test material. The introduction of the polymerase chain reaction (PCR) (SI) successfully overcame some of these disadvantages. 

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