The present investigation was aimed at understanding the molecular mechanism of gene amplification. Interplay of fragile sites in promoting gene amplification was also elucidated. The amplification promoting sequences were chosen from the Saccharomyces cerevisiae ARS, 5S rRNA regions of Plantago ovata and P. lagopus, proposed sites of replication pausing at Ste20 gene locus of S. cerevisiae, an...

Blood samples have traditionally been used as the main source of DNA for genetic analysis. However, this source can be difficult in terms of collection, transportation, and long-term storage. In this study, we investigated whether human nail clippings could be used as a source of DNA for SNP genotyping, null-allele detection, and whole-genome amplification. From extracted nail DNA, we achieved ...

  Background and Objective: Formalin-fixed paraffin-embedded tissues are a valuable source of DNA for molecular studies. We designed and optimized an efficient procedure for DNA extraction from formalin-fixed paraffin embedded tissues. Materials and Methods: Seventy three blocks of cervical paraffin-embedded tissues were investigated. DNA was extracted using 45 minutes boiling in alkaline sol...

DNA amplification by the polymerase chain reaction (PCR) was used to detect DNA of the Lyme disease spirochaete Borrelia burgdorferi. Primers that specify the amplification of a 145 basepair DNA fragment of the OspA gene of B. burgdorferi were used. The amplification product was detected by gel electrophoresis and ethidium bromide staining or by hybridization to a radiolabelled oligonucleotide ...

DNA amplification is essential to most nucleic acid testing strategies, but established techniques require sophisticated equipment or complex experimental procedures, and their uptake outside specialised laboratories has been limited. Our novel approach, recombinase polymerase amplification (RPA), couples isothermal recombinase-driven primer targeting of template material with strand-displaceme...

Chromosomal aberrations and dihydrofolate reductase gene amplification are observed in L5178Y mouse lymphoma cells after treatment with hydroxyurea. The types of aberrations include polyploidy, endoreduplication, chromosome fragmentation, and the presence of extrachromosomal DNA. Hydroxyurea-treated cells analyzed by cell sorting showed a subpopulation of cells with increased DNA and increased ...

In “Dynamics and Control of DNA Sequence Amplification”, we have formulated the PCR optimal control problem and discussed the strategies for solving it. In this chapter, we implement those strategies and solve a simple PCR optimal control problem. The foundational concepts of Optimal Control were developed as early as the 1750s in classical mechanics as the principle of least action, a variatio...

By using appropriate amounts of four bacteriophage phi 29 DNA replication proteins--terminal protein, DNA polymerase, protein p6 (double-stranded DNA-binding protein), and protein p5 (single-stranded DNA-binding protein)--it has been possible to amplify limited amounts of the 19,285-bp-long phi 29 DNA molecule by three orders of magnitude after 1 hr of incubation at 30 degrees C. Moreover, the ...

To analyze mutation in tomato genomic DNA by RAPD technology, certain factors influencing RAPD amplification, such as, DNA template, Taq DNA polymerase, Mg, dNTPs and primer, were studied to optimize RAPD amplification reaction system. RAPD amplification of tomato genomic DNA after low energy ion implantation along with soybean DNA was successfully performed. Compared to the control, genomic DN...

The application of microfluidic devices for DNA amplification has recently been extensively studied. Here, we review the important development of microfluidic polymerase chain reaction (PCR) devices and discuss the underlying physical principles for the optimal design and operation of the device. In particular, we focus on continuous-flow microfluidic PCR on-chip, which can be readily implement...