SELEX with Affinity Chromatography on RNA-RNA Interaction

RNA Affinity Chromatography

Affinity chromatography with an immobilized RNA enzyme.

M1 RNA, the catalytic subunit of Escherichia coli RNase P, has been covalently linked at its 3' terminus to agarose beads. Unlike M1 RNA, which is active in solution in the absence of the protein component (C5) of RNase P, the RNA linked to the beads is active only in the presence of C5 protein. Affinity chromatography of crude extracts of E. coli on a column prepared from the beads to which th...

Affinity chromatography of RNA-dependent DNA polymerase from RNA tumor viruses on a solid phase immunoadsorbent.

A solid phase immunoadsorbent specific for RNA-dependent DNA polymerase from murine and feline RNA tumor viruses has been prepared. The enzymes from murine and feline virus but not from avian virus bind to columns of this material. Bound enzymes can be eluted in active form. This method has permitted selective purification of viral enzyme from crude extracts of virus-transformed cells, since th...

RAPID-SELEX for RNA Aptamers

Aptamers are high-affinity ligands selected from DNA or RNA libraries via SELEX, a repetitive in vitro process of sequential selection and amplification steps. RNA SELEX is more complicated than DNA SELEX because of the additional transcription and reverse transcription steps. Here, we report a new selection scheme, RAPID-SELEX (RNA Aptamer Isolation via Dual-cycles SELEX), that simplifies this...

SELEX-on-a-Chip: Microchip Integration of the Sol-Gel Derived Affinity Column for Monitoring RNA-Protein Interaction

A microfluidic chip that integrates a sol-gel derived affinity column with a microheater is developed. This microchip includes five electric heaters covered with patterned PMMA for individual heat elution from five sol-gels with target molecules. This study is the first to demonstrate the detection of proteinaptamer interactions on the sol-gel microfluidic system and further describes possible ...