Automated purification and suspension array detection of 16S rRNA from soil and sediment extracts by using tunable surface microparticles.

ABSTRACT: Autonomous, field-deployable molecular detection systems require seamless integration of complex biochemical solutions and physical or mechanical processing steps. In an attempt to simplify the fluidic requirements for integrated biodetection systems, we used tunable surface microparticles both as an rRNA affinity purification resin in a renewable microcolumn sample preparation system and as the sensor surface in a flow cytometer detector. The tunable surface detection limits in both low- and high-salt buffers were 1 ng of total RNA ( approximately 10(4) cell equivalents) in 15-min test tube hybridizations and 10 ng of total RNA ( approximately 10(5) cell equivalents) in hybridizations with the automated system (30-s contact time). RNA fragmentation was essential for achieving tunable surf

SUBMITTER: Chandler DP 

PROVIDER: S-EPMC404419 | biostudies-literature | 2004 May

REPOSITORIES: biostudies-literature