BE/GEN-EN: Who's Doing What in a Complex Bioreactor? Stable Isotope Probing of Specific Degraders in Engineered Biological Treatment Processes

0221836 Aitken将使用稳定同位素方法，在完全封闭的浆液好氧生物反应器中，通过混合培养物和特定C-13标记的PAH底物的孵育制备重链DNA。 生物反应器配有气相色谱仪，可进行在线呼吸测量。 将从浆液中提取DNA和rRNA，通过PCR扩增，并在琼脂糖凝胶上分析。 比较rDNA（PCR）、rRNA（RT-PCR）和C-13标记的rDNA和rRNA的DGGE图谱，可以了解群落中代谢活跃的成员。 对选定的DGGE条带进行测序，并提交使用PHYLIP软件进行系统发育分析。

0221836 Aitken A stable isotope method will be used to prepare heavy DNA from incubations of mixed cultures and specific C-13 labelled PAH substrates in completely closed slurry aerobic bioreactors. The bioreactor is fitted with a gas chromatograph which allows on- line respirometric measurements. DNA and rRNA will be extracted from the slurry, amplified by PCR, and analyzed on agarose gels. Comparisons between the DGGE profiles obtainedfrom rDNA (PCR), rRNA (RT-PCR) and C-13 labelled rDNA and rRNA will provide insight into the metabolically active members of the community. Selected DGGE bands will be sequenced and submitted for phylogenetic analysis using PHYLIP software.