Amplification of source DNA is a nearly universal requirement for molecular biology applications. The primary methods currently available to researchers are limited to in vivo amplification in Escherichia coli hosts and the polymerase chain reaction. Rolling-circle DNA replication is a well-known method for synthesis of phage genomes and recently has been applied as rolling circle amplification...

The polymerase chain reaction (PCR) is the most widely used technique for the study of DNA. Applications for PCR have been extended significantly by the development of "long" PCR, a technique that makes it possible to amplify DNA fragments up to 40 kb in length. This article describes two novel applications of the long PCR technique, one which simplifies restriction mapping and another which en...

A modified in-gel DNA renaturation technique, which detects DNA sequences amplified greater than 7-fold in human DNA, was used to analyze gene amplification in surgical specimens of primary and metastatic ovarian carcinomas. Amplified DNA sequences were detected in two of eight tumors. Hybridization of these samples with different oncogene probes revealed that both tumors contained an amplified...

We report here an alternative and easy approach to generate sufficient quantity of minimal gene expression cassette (promoter–open reading frame–terminator) DNA by PCR amplification using high fidelity DNA polymerases. Isolating the minimal gene cassette DNA in large quantities by restriction digestion and gel extraction for biolistic-mediated ‘clean DNA’ transformation, is not only a cumbersom...

As previously reported, a novel low temperature (LoTemp) polymerase chain reaction (PCR) catalyzed by a moderately heat-resistant (MHR) DNA polymerase with a chemical-assisted denaturation temperature set at 85 °C instead of the conventional 94-96 °C can achieve high-fidelity DNA amplification of a target DNA, even after up to 120 PCR thermal cycles. Furthermore, such accurate amplification is ...

Rolling circle amplification (RCA) catalyzed by φ29 DNA polymerase offers a simple method for DNA amplification in the presence of a circular DNA template and its complimentary primer. RCA continuously produces long single-strand DNA using the strand displacement activity of polymerase during DNA synthesis. This property allows one to monitor the progress of a reaction by means of electrophores...

Different amplification protocols were evaluated for use with primer-extension preamplification (PEP). We hypothesized that a protocol known to improve amplification of long DNA fragments would improve efficacy of PEP. Eight DNA samples were preamplified by PCR using different protocols. Treatments consisted of the use of Taq DNA polymerase (T), Taq plus a second polymerase obtained from Pyroco...

A Loop-Mediated Isothermal Amplification (LAMP) assay was developed for the detection of pine pathogen Dothistroma septosporum (G. Dorog.) M. Morelet. The specificity LAMP tested using a selection needle fungi, including pini Hulbary, and Lecanosticta acicola (Thüm.) Syd.; only D. DNA amplified by test. In terms sensitivity, able to detect as little 1 pg total DNA. This enables extracted from d...

BACKGROUND Polymerase chain reaction (PCR) is one of the most important developments in modern biotechnology. However, PCR is known to introduce biases, especially during multiplex reactions. Recent studies have implicated the DNA polymerase as the primary source of bias, particularly initiation of polymerization on the template strand. In our study, amplification from a synthetic library conta...