An efficient, PCR based method for the selective amplification of DNA target sequences that differs by a single base pair is described. The method utilises the high affinity and specificity of PNA for their complementary nucleic acids and that PNA cannot function as primers for DNA polymerases.
1Department of Immunology, Waiter Reed Army Institute of Research, Washington, D.C. 20307-5100; 2Tropical Disease Unit, Division of Infectious Diseases, The Toronto Hospital and The University of Toronto, Toronto, Ontario, Canada M5G 2C4 In vitro transcription and translation are powerful tools for examining the s t ructure-funct ion relationships of proteins. Genes can be cloned into plasmid v...
Natalia E. Broude,
Sjaak van Heusden,
We present a protocol for effi cient amplifi cation of a large number of DNA targets using a single-tube polymerase chain reaction (PCR) based on PCR suppression. This method allows amplifi cation of each DNA target with only one target-specifi c primer whereas the other primer is common for all targets. Thus, this approach requires n+1 primers for n targets instead of 2n in conventional PCR an...
P E Varaldo,
I n the last few years, molecular hybridization methods have been used extensively in basic and applied virology because of their technical flexibility and high specificity. Using these techniques, the detection of DNA and RNA viruses directly from clinical specimens, the analysis of the specific transcriptional activity of viral genes in vitro and in vivo, and the study of virus-host relations...
In this study, two PCR-based methods (MSP-PCR and PCR-MP) were compared for their abilities to identify intraspecies variations of 23 isolates of Trichophyton rubrum, 78 isolates of Trichophyton interdigitale and 22 isolates of Microsporum canis, obtained mainly from patients in Lódź city. The results allowed to distinguish four types (containing two subtypes) characteristic for T. interdigital...
:Proceedings of the National Academy of Sciences of the United States of America1999
K W Kinzler,
The identification of predefined mutations expected to be present in a minor fraction of a cell population is important for a variety of basic research and clinical applications. Here, we describe an approach for transforming the exponential, analog nature of the PCR into a linear, digital signal suitable for this purpose. Single molecules are isolated by dilution and individually amplified by ...
We are very pleased to be able to bring you this special issue of iomolecular Detection and Quantification which focuses on digital CR (dPCR). The underpinning method of dPCR, which was coined n 1999 , actually predates qPCR  and is a powerful technique hat could offer improved sensitivity, precision and reproducibility 3]. Now is an exciting time for this method and there are numerous xa...
In this project, real time polymerase chain reaction (PCR) was utilized to study the mechanism of PCR inhibition through examination of the effect of amplicon length, melting temperature, and sequence. Specifically designed primers with three different amplicon lengths and three different melting temperatures were used to target a single homozygous allele in the HUMTH01 locus. The effect on amp...
We describe a method for producing specific PCR primers directly from PCR product, bypassing the usual need to know the primer sequence. Lack of abundance of primers derived from a PCR product is compensated for by the incorporation of an arbitrary 5'TAG sequence which acts as a surrogate template target for the bulk amplification phase. We use the technique to amplify clonospecific rearranged ...
INTRODUCTIONPCR-based whole-genome amplification (WGA) has the goal of generating microgram quantities of genome-representative DNA from picogram or nanogram amounts of starting material. This amplification should introduce little, or ideally no, representational bias. Unlike other techniques for WGA, PCR-based methods are generally less affected by DNA quality and are more applicable to DNA ex...