11:680:390 Lecture Notes - Lecture 14: Acridine, Nitrification, Lithotroph

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Culture-independent analyses: most microbes in nature cannot be cultured , enrichments and isolations disrupt microbial interactions, staining methods. Nonspecific all cells (living or dead) will be stained. Dyes that intercalate into nucleic acids (most commonly used: dapi, acridine orange) Combination of dyes that stain all cells and those that stain only live cells. Limited use with environment al samples: nitrification. Cell tags: gfp expressed constitutively) reveal the location of the bacterium in an environmental sample. Reporter genes: (gfp expressed conditionally) inform the presence of a specific compound in the environment. Small oligonucleotide probes that are homologous to 16s rrna genes. Identify organisms that contain a homologue of clone. Dna in a mixture; helps us answer who is there? : microbial community analysis based on pcr (environmental clone libraries), dna sequencing, phylogenetic analysis, how d o clone libraries work, collect microbial biomass from target environment and extract. Dna: amplify 16s rrna gene (separation based on size) c)

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