High-purity preparation of a large DNA dumbbell.

Efficient production of superior dumbbell-shaped DNA minimal vectors for small hairpin RNA expression

Genetic therapy holds great promise for the treatment of inherited or acquired genetic diseases; however, its breakthrough is hampered by the lack of suitable gene delivery systems. Dumbbell-shaped DNA minimal vectors represent an attractive, safe alternative to the commonly used viral vectors which are fraught with risk, but dumbbell generation appears to be costly and time-consuming. We devel...

A simplified procedure for large scale preparation of purified animal DNA-dependent RNA polymerase.

A procedure is described for obtaining DNA-dependent RNA polymerase form B from calf thymus in high purity. The procedure includes homogenization in low salt, sedimentation of chromatin at 50 000 x g, adsorption to DEAE-cellulose in a batch procedure, DNA-agarose chromato­ graphy, and chromatography on DEAE-Sephadex. Analyses of the preparation by band sedimenta­ tion and gel-electrophoresis in...

Preparation of High-Purity Grade Ammonium Sulfate from Locally Available Products

Preparation of factor VII concentrate using CNBr-activated

  Background: Factor VII concentrates are used in patients with congenital or acquired factor VII deficiency or treatment of hemophilia patients with inhibitors. In this research, immunoaffinity chromatography was used to purify factor VII from prothrombin complex (Prothrombin-Proconvertin-Stuart Factor-Antihemophilic Factor B or PPSB) which contains coagulation factors II, VII, IX and X. The a...

Improved preparation and identification of aristolochic acid-DNA adducts by solid-phase extraction with liquid chromatography-tandem mass spectrometry.

Aristolochic acid (AA) is a known nephrotoxin and potential carcinogen, which can form covalent DNA adducts after metabolic activation in vivo and in vitro. A simple method for preparation and characterization of aristolochic acid-DNA adducts was developed. Four AA-adducts were synthesized by a direct reaction of AAI/AAII with 2'-deoxynucleosides. The reaction mixture was first cleaned-up and p...