Applications of the polymerase chain reaction to genome analysis

Abstract

The objectives of the Human Genome Project are to create high-resolution genetic and physical maps, and ultimately to determine the complete nucleotide sequence of the human genome. The result of this initiative will be to localize the estimated 50,000-100,000 human genes, and acquire information that will enable development of a better understanding of the relationship between genome structure and function. To achieve these goals, new methodologies that provide more rapid, efficient, and cost effective means of genomic analysis will be required. From both conceptual and practical perspectives, the polymerase chain reaction (PCR) represents a fundamental technology for genome mapping and sequencing. The availability of PCR has allowed definition of a technically credible form that the final composite map of the human genome will take, as described in the sequence-tagged site proposal. Moreover, applications of PCR have provided efficient approaches for identifying, isolating, mapping, and sequencing DNA, many of which are amenable to automation. The versatility and power provided by PCR have encouraged its involvement in almost every aspect of human genome research, with new applications of PCR being developed on a continual basis.

PCR-based microsatellite polymorphisms in the detection of loss of heterozygosity in fresh and archival tumour tissue

“…This group of DNA polymorphisms have collectively been named microsatellites (Litt & Luty, 1989). Microsatellite markers have already proved to be very useful in mapping of several genetic disease genes (Hanzlik et al, 1990;Wijmenga et al, 1990;Bell et al, 1991;CannonAlbright et al, 1992) and genome mapping (Jabs et al, 1991;Rose, 1991). Microsatellite markers show a number of obvious advantages over Southern blot approaches as they (i) can be detected by PCR, (ii) are not restricted to telomeric or centromeric regions, (iii) are multi-allelic, (iv) can be used in a multiplex PCR (multiple primer sets per PCR reaction) and (v) need a very small amount of input DNA.…”

Archaeal DNA polymerases in biotechnology

“…When a gene is inserted into a vector, new restriction sites are needed at the DNA ends, which can be also NP not published engineered by PCR. Moreover, PCR is an efficient approach for identifying, isolating, mapping, and sequencing DNA, and many of these techniques are used in human genome research (Rose 1991). By using a PCR primer that is specific to DNA from a pathogen or to a mutant site in the human genome, diagnostic PCR has many applications in identifying mutant alleles, disease causing pathogens and cancer research (Emmanuel 1993;McCormick 1989).…”

Determination of avian endogenous provirus ‐ cellular junction sequences using inverse polymerase chain reactions

“…Moreover, consistently satisfactory restriction fragment polymorphisms (RFLPs) may be difficult to obtain. The development of the polymerase chain reaction (PCR) wherein target DNA sequences are amplified exponentially has found wide use in genetic analysis (Rose, 1991).…”