Department of Agronomy and Range Science, University of California, Davis, California 95616-8515 Recent advances in PCR have made this technique one of the most powerful tools for a wide spectrum of molecular analyses, such as genome mapping, molecular evolution, diagnosis of genetic disease, and forensic sciences. ~ Many PCR applications involve the specific and reproducible amplif icat ion of...

The polymerase chain reaction (PCR) has recently evolved as a standard laboratory technique, popular in all areas of molecular biology research. However, the technique still has two limitations: the relatively low fidelity of Taq polymerase when compared with other polymerases (1), and its inability to efficiently amplify fragments higher than 3 kbp (2, 3). Although these two issues are irrelev...

DNA microarray-based screening and diagnostic technologies have long promised comprehensive testing capabilities. However, the potential of these powerful tools has been limited by front-end target-specific nucleic acid amplification. Despite the sensitivity and specificity associated with PCR amplification, the inherent bias and limited throughput of this approach constrain the principal benef...

Traditional asymmetric PCR uses conventional PCR primers at unequal concentrations to generate single-stranded DNA. This method, however, is difficult to optimize, often inefficient, and tends to promote nonspecific amplification. An alternative approach, Linear-After-The-Exponential (LATE)-PCR, solves these problems by using primer pairs deliberately designed for use at unequal concentrations....

PCR is an indispensable technique used in DNA analysis. However, with the traditional methods, the time spent on amplification and the subsequent analysis of PCR products is generally long. Therefore, it is essential to improve these two steps so that the whole procedure can be made faster. In the present work, with lambda-DNA as the control template, the amplification of 300-bp fragment could ...

Whole-genome amplification (WGA) techniques are used for non-specific amplification of low-copy number DNA, and especially for single-cell genome and transcriptome amplification. There are a number of WGA methods that have been developed over the years. One example is degenerate oligonucleotide-primed PCR (DOP-PCR), which is a very simple, fast and inexpensive WGA technique. Although DOP-PCR ha...

We present two examples of exponential nucleic acid amplification with the polymerase chain reaction (PCR) in the presence of only one amplification primer. Cloning and sequencing of the PCR products generated by amplification of human genomic DNA revealed that the amplified sequence contained only one primer and its complement, at the two ends of the PCR product. Although these experiments wer...

In light of the strict legal scrutiny surrounding DNA typing at this time, it has become necessary to systematically address the issue of PCR contamination. To precisely define the parameters affecting PCR contamination under casework analysis conditions, PCR amplification reactions were intentionally compromised by employing sub-standard laboratory technique and by introducing secondary source...

Quantitative applications of the Polymerase Chain Reaction (PCR), also known as Quantitative-PCR (Q-PCR) are intended either to determine the number of copies of a given nucleic acid sequence, or more generally, to determine the relative abundance of two sequences. Current methods to determine exact numbers of molecules overcome the determination of the amplification rate by assuming identical ...

An efficient cDNA amplification procedure is described for determining of the 5' and 3' ends of mRNAs and cloning full-length cDNAs. In this approach, a double-stranded (ds) adaptor is ligated to both ends of a library of ds cDNA by T4 DNA ligase. This adaptor-ligated ds cDNA is then used to selectively amplify 5'- or 3'-cDNA fragments by PCR with a combination of gene-specific and adaptor-spec...