Plasmid purification is a basic tool of molecular biologists. Although the development of plasmid isolation kits utilizing silica spin columns reduced the time and labor spent on plasmid purification, achieving large plasmid DNA yields still requires significant time and effort. Here we introduce the Miraprep, a rapid protocol that allows isolation of plasmid DNA using commercial Miniprep kits,...

A simple method for the isolation of pure and high-yield DNA from whole blood, suitable for restriction enzyme digestion, is described. The steps of the procedure are as follows: cell lysis with NH4Cl, NaHCO3, EDTA; digestion with proteinase K in the presence of SDS; extraction with phenol-chloroform-isoamyl alcohol; and precipitation with ethanol. The 260 nm/280 nm absorbance ratio showed a me...

In plants, secondary metabolites and polysaccharides interfere with genomic isolation procedures and downstream reactions such as restriction enzyme analysis and gene amplification. The removal of such contaminants needs complicated and time-consuming protocols. In this study, a simple, rapid and efficient method for leaf DNA extraction was optimized. This method use small amount of plant mater...

OBJECTIVE To compare direct and indirect bead-based T-cell isolation followed by magnetic bead-based DNA isolation. METHODS T-cells were isolated by direct or indirect selection with magnetic bead coated antbiodies followed by magnetic bead-based automated DNA isolation in 10 healthy subjects. Purity of T-cells, purity of DNA (by A260/A280 ratio measurement) and DNA concentration were assesse...

We describe a rapid, simple, and reliable procedure for routine isolation of Mycobacterium leprae DNA from slit-skin swab specimens. This one-step DNA extraction method is based on the utilization of ChelexR 100, a chelating ion exchange resin. Slit-skin swab specimens from seven untreated leprosy patients at Baba Baghi Leprosy Hospital in Iran were processed by this procedure. The polymer...

The efficiency of one-step and multi-step protocols of DNA isolation from lysed sputum samples containing the Mycobacterium tuberculosis complex has been compared. DNA was isolated using spin-cartridges containing a special silica-based sorbent modified with fluoroplast and polyaniline, or using an automated isolation system. One-step isolation using the obtained sorbent has been shown to ensur...

The minipreparation of plasmid DNA, or miniprep, is a small-scale isolation of plasmid DNA from bacteria. Two main approaches have been used for this isolation; the first and more traditional technique is often a variation on the original protocol by Birnboim and Doly (1979). The second uses a special silica matrix to bind DNA and then release it under certain conditions. The matrix is often in...

INTRODUCTION In HIV-infected patients on combination antiretroviral therapy (cART), the detection of episomal HIV 2-LTR circles is a potential marker for ongoing viral replication. Quantification of 2-LTR circles is based on quantitative PCR or more recently on digital PCR assessment, but is hampered due to its low abundance. Sample pre-PCR processing is a critical step for 2-LTR circles quanti...

Aim. Investigate the biological properties of actinomycetes from rhizosphere Colobanthus quitensis (Kunth) Bartl. and study their ability to produce bioactive compounds. Methods. Microbiological (isolation, synthesis compounds), genetic engineering (isolation analysis total DNA, DNA gel electrophoresis, polymerase chain reaction, sequencing), bioinformatic (phylogenetic analysis) methods. Resul...