Molecular Monitoring of Microbial Diversity in Anaerobic Wastewater Treatment Systems with an Identification Array and Start-up of ID-array platform

Kees Roest, Hans G.H.J. Heilig, Willem M. de Vos, Alfons J.M. Stams, Antoon D.L. Akkermans.

Laboratory of Microbiology, Wageningen University, Hesselink van Suchtelenweg 4, NL-6703CT WAGENINGEN, Nederland, Phone: +31-317-483118 / 484250, Fax: +31-317-483829

We started with the development of a DNA macro array for detection of molecular diversity in Upflow Anaerobic Sludge Blanket (UASB) sludge. Approximately 80-mer variable regions of cloned and identified 16S rDNA sequences from sludge, were amplified and blotted onto a filter. This approach was evaluated by performing a reverse hybridisation with labelled 16S rDNA from a selection of the initial clones. Focus was on the V6 region, which appeared to be promising for identification with an array although cross-hybridisation remained a problem. A major source for this cross-hybridisation was the presence of the primer sites, which are highly conserved. To circumvent this, primer sites of PCR products were removed by enzymatic action. More variable regions of sequenced clones will be used for the development of a multiple probe array. The developed macro array will be miniaturised to an identification DNA micro array and used for rapid molecular detection of microbial diversity in sludge of wastewater treatment samples.

Furthermore, we initiated the start-up of an ID-array platform. This platform should be informal and informative. We can notify each other about interesting seminars, papers, vacancies, etc. Therefore I'm setting up a mailing list and a web page. I already have several addresses, however I'd like to expand the mailing list with more people working with identification DNA arrays. 
          The aim of the web page is to give a good overview of the research with identification DNA arrays in the Netherlands. Therefore I'd like to receive short descriptions of research (plans) dealing with identification arrays, including affiliation and any other relevant information (e.g. available array equipment). Furthermore suggestions for the web page (e.g. links) or the 'ID-array platform' in general are welcome.