Search results for "Agar gel"

Sliding-end-labelling

1986

Abstract A method, termed ‘sliding-end-labelling’, has been devised to avoid a frequent artifact in nucleosome positioning by indirect end labelling, namely the appearing of DNA fragments originated by two nuclease cuts, one of them lying within the region covered by the probe. The method is applied to the nucleosome positioning in the yeast SUC2 gene for invertase.

Stability of PEI–DNA and DOTAP–DNA complexes: effect of alkaline pH, heparin and serum

2001

Abstract DNA complexes formed with nonviral vectors such as polyethylenimine (PEI) or 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) are widely used in gene therapy. These complexes prevent the interaction of DNA with the fluorescent probes usually employed to quantify DNA. We thus studied the procedures for DNA quantification from DNA complexes as well as their stability in the presence of DNase or mouse, rat and human sera. Release of the DNA from its complexes was accomplished by increasing the pH of the medium (from 7.3 to 13.4) or by adding heparin. The stability against degradation was tested in vitro, by incubating the complexes at 37°C in the presence of DNase I and sera from the …

Antibacterial studies, DNA oxidative cleavage, and crystal structures of Cu(II) and Co(II) complexes with two quinolone family members, ciprofloxacin…

2005

Nine coordination compounds of Cu(II) and Co(II) with Ciprofloxacin (HCp) and Enoxacin (HEx) as ligands have been prepared and characterized. Single crystal structural determinations of [Cu(HCp)2(ClO4)2].6H2O (1) and [Co(HEx)2(Ex)]Cl.2CH(3)OH.12H2O (4) are reported. The crystal of 1 is composed of [Cu(HCp)2(ClO4)2] units with the two perchlorate anions semicoordinated, and uncoordinated water molecules. The copper ion, at a crystallographic inversion centre, is in a tetragonally distorted octahedral environment. The structure of 4 consists of cationic monomeric [Co(HEx)2(Ex)]+ units, chloride anions, and uncoordinated methanol and water molecules. The complex is six-coordinate, with a sligh…

MboII endonuclease heat inactivation before agarose gel electrophoresis to prevent artifactual bands in restriction patterns

1999

Orthogonal electrophoretic fractionation of DNA in agarose gels.

2008

We developed an electrophoretic procedure, using Voltage Gradient Gel Electrophoresis (VGGE), which allows to obtain both an improvement of the resolution power of the system in orthogonal fractionation of DNA and, mainly, an about fourfold enhancement of hybridization signals in Southern blotting applications.

Voltage gradient electrophoresis of nucleic acids on agarose gels.

1993

A very simple method is described which allows the separation of DNA molecules in a wide molecular weight range (from 0.6 to about 30 kb) in the same electrophoresis agarose gel. This is based on the achievement of a voltage gradient through a simple device consisting of a Plexiglas plate placed slantwise with respect to the gel surface plane, submerged in the electrophoretic running buffer. Further applications of our system are also described.

Enhanced hybridization labeling signals in Southern blotted DNAs fractionated with voltage gradient gel electrophoresis.

1998

An enhancement of hybridization labeling signals is demonstrated in Southern blotted DNAs, fractionated by voltage gradient gel electrophoresis. This enhancement is due to a reduced thickness of each single nucleic acid band in the gel as a consequence of the gradient effect, corresponding to an increased concentration of DNA per unit area.

Genetic transfer of the mcd gene in soil.

2003

Aims: To investigate the role of horizontal gene transfer of mcd (methylcarbamate-degrading) gene in high genetic diversity of carbofuran-degrading bacteria. Methods and Results: The actuality of genetic transfer from degraders to an Agrobacterium tumefaciens strain was determined in liquid medium. The mcd gene was chosen for transfer experiments. Transconjugants were obtained irrespective of the type of the donor strain (Gram-positive or Gram-negative), size of the inoculum, or nature and concentration of the pesticide in the medium. Soil microcosms, inoculated with or without the donor and/or recipient strains were used. The size of the initial degrading population (treated or untreated s…

A report of an international collaborative experiment to demonstrate the uniformity obtainable using DNA profiling techniques

1992

This paper describes a collaborative exercise intended to demonstrate whether uniformity of DNA profile results could be achieved between different European laboratories. It was shown that this goal can be obtained provided that a common protocol is followed (specifically the use of a common electrophoretic buffer as being the most important parameter). Generally, lower molecular weight loci (with lower molecular weight fragments) such as YNH24 perform better than higher molecular weight loci such as MS43a. The results of the exercise are discussed in relation to the objectives of the European DNA profiling group (EDNAP).

Pulsed field gel electrophoresis and genome size estimates

2015

Pulsed field gel electrophoresis (PFGE) is a quick and reliable procedure to resolve DNA molecules larger than 30 kb by applying an electric field that periodically changes direction. This technique can be used to estimate genome size of a microorganism, to reveal if a genome is circular or linear, to indicate the presence of megaplasmids, and to show if a strain contains only one or more chromosomes.