Date post: | 22-Dec-2015 |
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Molecular Microbiology
Genetic Recombination
• Vertical – Binary fission– Budding
• Horizontal– Conjugation– Transduction– Transformation
Conjugation
Transduction
Transformation
Griffith Experiment
Biotechnology
• Recombinant DNA• Genetic Engineering• Disease treatments
– SCID– SSA– CF
• Make products– Vaccines– Enzymes– Insulin– Hormones
Plasmids
• DNA• Replicate
independently• Genetic Vectors
– Small– Stable– Marker inserted– Gene expression
Background
• Donor• Recipient• Competent Cell
– Natural• Change cell wall• Change PM
– Artificial• Temperature• Salts (CaCl2)
• Marker• Cloning Site
LB Broth and Agar
• Lysogeny Broth– Luria Broth– Luria-Bertani
• Nutritionally Rich– Peptides, casein– Vitamins– Minerals (NaCl)– Trace elements
• Biotech– Cultivate E.coli
Procedure• Follow lab manual!!• Reader
– Directs– Keeps time
• Materials– LB plates
• 2 types– LB– LB-amp
• 2 situations– Before– After
• Label, Label, Label– pGreen plasmid– CaCl2– Micropipets – Spreader– Loop inoculation
• Temperature– Incubator– Baths
• Ice• Heat
Results
• Marker– Fluorescence
• Cloning site– R-ampicillin
• Colony observation– Natural light– UV light
• Calculate transformation efficiency
Questions?
Glow Fish