EEMB 3 Lecture Notes - Lecture 6: 16S Ribosomal Rna, Kary Mullis, Craig Venter
21 Apr 2019
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Kary mullis - nobel prize in chemistry 1993. Taq polymerase discovered by thomas brock & hudson freeze. Polymerase that is found in high temperatures thermally stable. Isolation of microbes from hot spring source of thermally stable polymerase (taq) used in pcr. A way to isolate molecules from a mixture of molecules. Cultivation independent techniques do not need to culture microbes. Can look at sequence of genes to understand diversity. Lyse cells, extract dna, amplify 16s rdna (pcr) Used engineered plasmids to insert specific genes (ligating) Adding 16s gene causes the lac z gene to ligate (split) so that it no longer produces b-galactosidase. Insert plasmid into e. coli transformation (ability of a microbe to take up extracellular. Pour transformed culture onto plates that contain antibiotics on it. Organisms without antibiotic resistance won"t live on that plate. Organisms that successfully transformed plasmids will grow on petri dish.
