observation hypothesis experimental design (including methods) results inference camp wildness 2004...
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Observation
Hypothesis
Experimental Design (including Methods)
Results
Inference
Camp Wildness 2004 Ward Lab Research Project
The concept of housekeeping genes
ATPase
DNA-dependantDNA polymerase
DNA-dependantRNA polymerase
ribosomecomponents
Ribosome components
Small subunit (16S) rRNA
Highly variablesegment
Highly conservedsegment
PCR amplification of SSU rRNA genes
Focus on a particular gene in the genome
Extract DNA
PCR
Mixed 16S rRNAgenes
Molecular analysisof microbial communities
PCR amplificationOf 16S rRNA gene
Agarose gel analysis of PCR product
Separate by DGGE
Increasing D
enaturant
Mixed Population DNAs
+16S rRNA Gene G+C “Clamp”
PCR Primers G+C-Tailed ProductSeparate onDenaturingGradient
Gel
PCR Amplification
Denaturing Gradient Gel Electrophoresis
Separation Based onDifferences in
Nucleotide Sequence (G+C content) and Melting
Characteristics
B CMix A
Purified Bands for Sequence Analysis
DGGE analysis of 16S rRNA sequences
DGGE study of temperature distribution of Octopus Springcyanobacterial mat 16S rRNA variants
ecotype
Extract DNA
PCR
Native microbialpopulations
Mixed 16S rRNA
Separate by DGGE
Sequencing
Analyzing distributionof molecular diversity
Denaturing GradientGel Electrophoresis
Phylogenetic identification
Gloeobacter violaceus
Pseudoanabaena galeata
OS Type P Isolate
Oscillatoria limnetica
Oscillatoria sp. 7515
Microcoleus sp.
OS C1 Isolate
Synechococcus sp. 6301
Oscillatoria sp. 6304
Oscillatoria amphigranulata
OS Type I
Spirulina sp.
Anabaena cylindrica
Pleurocapsa sp.
Synechocystis sp.
Phormidium ectocarpi
OS C9 Isolate
0.01
OS Type J
OS Type B Isolate
OS Type B' Isolate
OS Type A''
OS Type A'
OS Type A
OS Type A'''
Yellowstone cyanobacterialmat cyanobacterial 16S rRNAdiversity
Readily cultured
predominant in situ
A natural view of microbiology
Observation
Hypothesis
Experimental Design (including Methods)
Results
Inference
Camp Wildness 2004 Ward Lab Research Project