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Observation Hypothesis Experimental Design (including Methods) Results Inference

Camp Wildness 2004 Ward Lab Research Project. Observation Hypothesis Experimental Design (including Methods) Results Inference. The concept of housekeeping genes. ATPase. DNA-dependant DNA polymerase. DNA-dependant RNA polymerase. ribosome components. Ribosome components.

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Observation Hypothesis Experimental Design (including Methods) Results Inference

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  1. Camp Wildness 2004 Ward Lab Research Project Observation Hypothesis Experimental Design (including Methods) Results Inference

  2. The concept of housekeeping genes ATPase DNA-dependant DNA polymerase DNA-dependant RNA polymerase ribosome components

  3. Ribosome components

  4. Highly variable segment Highly conserved segment Small subunit (16S) rRNA

  5. PCR amplification of SSU rRNA genes Focus on a particular gene in the genome

  6. Extract DNA PCR Mixed 16S rRNA genes Molecular analysis of microbial communities PCR amplification Of 16S rRNA gene Agarose gel analysis of PCR product

  7. Separate by DGGE

  8. PCR Amplification Mixed Population DNAs PCR Primers G+C-Tailed Product Separate on Denaturing Gradient Gel + 16S rRNA Gene G+C “Clamp” Denaturing Gradient Gel Electrophoresis Purified Bands for Sequence Analysis A B C Mix Separation Based on Differences in Nucleotide Sequence (G+C content) and Melting Characteristics DGGE analysis of 16S rRNA sequences Increasing Denaturant

  9. DGGE study of temperature distribution of Octopus Spring cyanobacterial mat 16S rRNA variants ecotype

  10. Extract DNA PCR Mixed 16S rRNA Separate by DGGE Phylogenetic identification Analyzing distribution of molecular diversity Denaturing Gradient Gel Electrophoresis Native microbial populations Sequencing

  11. Pleurocapsa sp. Phormidium ectocarpi Synechocystis sp. Anabaena cylindrica Spirulina sp. predominant in situ 0.01 OS Type I Oscillatoria amphigranulata OS C1 Isolate Synechococcus sp. 6301 Microcoleus sp. Oscillatoria sp. 7515 Oscillatoria limnetica Oscillatoria sp. 6304 OS Type P Isolate Pseudoanabaena galeata Gloeobacter violaceus OS Type J OS Type B' Isolate OS Type B Isolate OS Type A''' OS Type A'' OS Type A' OS Type A OS C9 Isolate Yellowstone cyanobacterial mat cyanobacterial 16S rRNA diversity Readily cultured

  12. A natural view of microbiology

  13. Camp Wildness 2004 Ward Lab Research Project Observation Hypothesis Experimental Design (including Methods) Results Inference

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