The DNA amplification performed by terminal protein-primed replication systems has not yet been developed for its general use to produce high amounts of DNA linked to terminal protein (TP). Here we present a method to amplify in vitro heterologous DNAs using the Φ29 DNA replication machinery and producing DNA with TP covalently attached to the 5' end. The amplification requires four Φ29 protein...

The reported genome-walking methods still suffer from some deficiencies, such as cumbersome experimental steps, short target amplicon, or deep background. Here, a simple and practical fork PCR was proposed for genome-walking. employs primer set of three random oligomers to implement walking task. In primary PCR, the low-stringency amplification cycle mediates binding places on genome, producing...

Single cell whole genome amplification is susceptible to amplification biases that impact the accuracy of single cell sequencing data. To address this, we have developed a microfluidic device for the isolation and purification of genomic DNA from individual cells. The device uses a micropillar array to physically capture single cells and its chromosomal DNA upon extraction. The extracted DNA is...

We describe conditions for rolling-circle amplification (RCA) of individual DNA molecules 5–7 kb in size by >109-fold, using 29 DNA polymerase. The principal difficulty with amplification of small amounts of template by RCA using 29 DNA polymerase is ‘‘background’’ DNA synthesis that usually occurs when template is omitted, or at low template concentrations. Reducing the reaction volume while k...

methylated and un-methylated control dna is an important part of dna methylation studies. although human and mouse dna methylation control sets are commercially available, in case of methylation studies on other species such as animals, plants, and bacteria, control sets need to be prepared. in this paper a simple method of generating methylated and un-methylated control dna is described. whole...

e-MagnetoMethyl IP is a new method for electrochemical analysis of global DNA methylation. It avoids bisulfite treatment, PCR amplification, and enzyme-based signal generation can detect differences as low 5% in methylation levels.

Identification of prognostic and predictive genomic markers requires long-term clinical follow-up of patients. Extraction of high-quality DNA from archived formalin-fixed, paraffin-embedded material is essential for such studies. Of particular importance is a robust reproducible method of whole genome amplification for small tissue samples. This is especially true for high-resolution analytical...

Detection of rare mutant alleles in an excess of wild type alleles is increasingly important in cancer diagnosis. Several methods for selective amplification of a mutant allele via the polymerase chain reaction (PCR) have been reported, but each of these methods has its own limitations. A common problem is that Taq DNA polymerase errors early during amplification generate false positive mutatio...

we investigated the use of dna amplification by polymerase chain reaction (per) for detection of mycobacterium tuberculosis in 300 patients who were suspected of having pulmonary tuberculosis and compared the results with culture results which were performed in parallel with pcr. two-thirds of each sample was processed for smear and culture by standard methods and one-third was prepared for dna...