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Molecular Genetics and Microbiology
Johanna Rommens

MGY376November 14 2011 On the way to a 16S rRNA gene sequence for your unknown1 Verify API identification of washroom sample by 16S sequencing 16Swhole gene highly conservedneed ribosomes rRNA Why 16S Not xS5S wouldnt have enough information to distinguishneed highly conserved regions but also need variable regions to distinguish different organisms23S is also fine but more to sequenceo All the database information is based on 16Sfor comparison purposesExtract DNA by freezing and thawing Treat sample with protinease K to rid of excess protein Amplify 16S using universal primers First half of moleculeo Colour coded to indicate degree of conservedness o Use more conserved sequences for PCR primers o Variable regions useful because distinguish btw families etc Use as molecular clock Run PCR product on agarose gel
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