Plasmid DNA chromatography is a powerful field in constant development and evolution. The use of this technique considered mandatory the production an efficient safe formulation to be applied for plasmid-mediated gene therapy. Concerning this, search ideal chromatographic support/ligand combination motivated scientist pursue continuous improvement on plasmid performance, looking progression lig...

Single stranded DNA (s.s.DNA) comprising 1-2% of the total nuclear DNA was isolated by an improved method of hydroxyapatite chromatography from native nuclear DNA3 of embryonic chick cells, labeled for several cell generations with 3H-thymidine. Small quantities of 3H-DNA were annealed with a large excess of unlabeled DNA or polysomal RNA from chick embryos. Hybridization kinetics (monitored by...

Column chromatography has evolved to provide a rapid and effective alternative to more laborious methods for preparing high-quality DNA, such as CsCl-gradient centrifugation. This unit describes the use of a column made of a unique anion-exchange resin that selectively binds nucleic acids, allowing rapid separation of DNA from contaminating RNA, proteins, carbohydrates, and metabolites. The pro...

Many of the most widely employed operations in molecular biology hinge upon the use of single-stranded DNA as a probe or template. Here we report a straightforward method by which to produce long single-stranded DNA molecules using the polymerase chain reaction (PCR) in combination with immobilized metal affinity chromatography (IMAC). We demonstrate that a tag consisting of six successive 6-hi...

The application of a 32P-postlabeling assay for 7-methylguanines in DNA was studied either by labeling the imidazole ring-opened dinucleotide derivatives or by using strong-anion-exchange column chromatography for the adduct enrichment from normal nucleotides. Data showed that 7-methylguanines can be efficiently labeled as dinucleotides when in vitro methylated DNA was first imidazole ring-open...

The affinity chromatography procedure described in this unit uses DNA containing specific recognition sites for the desired protein that has been covalently linked to a solid support. Preparation of a DNA affinity resin, including cyanogen bromide (CNBr) activation of the agarose support, is described, and an alternate protocol provides a method to couple DNA to commercially available CNBr-acti...

Denaturing High-Performance Liquid Chromatography (DHPLC) is a recently developed technique forthe detection of single nucleotide polymorphisms (SNPs) and mutations. It involves the comparisonbetween two or more DNAs as a mixture of denatured and reannealed PCR products. The methodologyis based on the principle of reversed phase liquid chromatography and uses a unique DNA sepa...