Affinity chromatography on columns containing DNA has been successfully used in the isolation and purification of DNA dependent polymerases and other DNA-binding proteins as well as for the purification of nucleic acids hybridizable to the column bound DNA [l] . Two drawbacks of DNA column materials used so far have limited their application: firstly, in the most widely used column materials DN...

We present here an improved RNA purification method using fast performance liquid chromatography (FPLC) size-exclusion chromatography in place of denaturing polyacrylamide gel electrophoresis (PAGE). The method allows preparation of milligram quantities of pure RNA in a single day. As RNA oligonucleotides behave differently from globular proteins in the size-exclusion column, we present standar...

Cellulose is a renewable and biodegradable natural biopolymersynthesized mainly in plants such as wood, bark, cotton fiber, green algae, some bacteria and animals. Pre-treatment processes such as alkaline purification and enzymatic purification are applied in order to create a proper sorption characteristic of regenerated cellulose fiber. Prior to the application of cellulose/cellulose derivati...

This chapter describes the purification and crystallization of oxygen-evolving photosystem II core dimer complex from a thermophilic cyanobacterium Thermosynechococcus vulcanus. Procedures used for purification of photosystem II from the cyanobacterium involves cultivation of cells, isolation of thylakoid membranes, purification of crude and pure photosystem II core complexes by detergent solub...

Small interfering RNA (siRNA)-induced gene silencing shows great promise in genomic research and therapeutic applications. siRNA duplexes are typically assembled from complementary synthetic oligonucleotides. High-purity single-stranded species are required for in vivo applications. Methods for separation, characterization, and purification of short RNA strands have been developed based on reve...

The conventional procedure for the purification of the high affinity K+ uptake ATPase (KdpABC) from Escherichia coli involves a tedious three-column protocol (final enzyme purity, approximately 90%; activity yield, 6.5% (Siebers, A., and Altendorf, K. (1988) Eur. J. Biochem. 178, 131-140)). We have now developed a highly effective one-column (Fractogel TSK AF-Red) protocol yielding an enzyme pr...

The strategies employed in chromatography steps play a key role in downstream processes for monoclonal antibody (mAb) manufacture. This work addresses the integrated optimisation of chromatography step sequencing and column sizing in mAb purification processes. Chromatography sequencing decisions include the resin selection at each typical step, while the column sizing decisions include the num...

A rapid and simple two-step procedure suitable for both small- and large-scale purification of pediocin-like bacteriocins and other cationic peptides has been developed. In the first step, the bacterial culture was applied directly on a cation-exchange column (1-ml cation exchanger per 100-ml cell culture). Bacteria and anionic compounds passed through the column, and cationic bacteriocins were...

A new passive solar water pasteurization system based on density difference flow principles has been designed, built and tested. The system contains no valves and regulates flow based on the density difference between two columns of water. The new system eliminates boiling problems encountered in previous designs. Boiling is undesirable because it may contaminate treated water. The system with ...

Protein constructs and purification For purification of NFκB:IRF-7:IRF-3, a freshly transformed colony was transferred into 10 ml LB broth containing 100 μg/ml ampicillin and grown overnight at 37°C to saturation. This overnight culture was used to inoculate 1 liter LB broth containing 100 μg/ml ampicillin and grown at 37°C to an OD600 of about 0.5. The culture was then induced with 0.4 mM isop...