Lab Exam II Microbiology Flashcards
Lyses cells
Sodium hydroxide/Sds
Precipitates debris
KOAc & detergent
Precipitates DNA
90% ethanol
Resuspend’s DNA
Triss EDTA buffer
Chelates calcium and magnesium
EDTA
Washes plasmid
70% ethanol
Can differentiate between endogenous and DNA that is not from the cell. Spares the bacterium from infection – recognizes palindromic sequences.sticky and allow them to combine with complementary single-stranded DNA. Add last.
Restriction endonucleases
nicks are annealed by…
DNA ligase
Small colonies that have not taken up the plasmid (White) that form around a large colony that has released beta lactamase which degrades ampicillin and it’s vicinity.
Satellite colonies
Degrades ampicillin
Beta lactamase
Helps us digest carbohydrates and synthesize certain vitamins. We provide E. coli with a nice warm place to live with food. It is also protected from viruses and competition with other bacteria.
E. coli
Is inhibiting the growth of one type of organism while permitting the growth of another as with an antibiotic.
Selection
When you are making a distinction between different organisms such as in a blue white screening.
Screen
Use reverse transcriptase to synthesize cDNA from post transcriptionally modified mRNA. The cDNA can be correctly translated in a prokaryotic cell.
How would you find a gene?
Extra chromosomal DNA commonly used as cloning vector. Origin of replication, selectable marker, cloning site into which the gene can be inserted.
Plasmid
Isolate DNA. Cleve DNA with restriction endonucleases and splice into a vector. Transform cells. Use selections and screens to elucidate which clones have taken up plasmid. Replica plating. Use of a primer will help you elucidate the clone expressing The gene of interest.
Making a gene library
Cleves media having x-gal so bacteria producing the enzyme turn blue.
There must be a recognition sequence in the beta-galactosidase Jean so that a gene of interest may be inserted and interrupt the beta galactosidase gene. Transformants will be white. Plate on antibiotic/X gal media.
Beta galactosidase
Bioxidation in which molecular oxygen can serve as the final electron acceptor.
Aerobic
Bioxidation in which organic ions other than oxygen such as nitrate or sulfate can serve as the final electron acceptor.
Anaerobic
Biooxidative process not requiring oxygen in which an organic substance serves as the final electron acceptor (lactic acid or alcohol fermentation. Yellow/yellow with gas)
Fermentation
Alkaline slant (red) and acid butt (yellow) with or without gas production breaks in auger butt.
Triple sugar iron test. Fermentation of glucose.
Acid slant yellow and acid butt (yellow) with or without gas production.
Triple sugar iron test. Lactose and or sucrose fermentation.
Alkaline slant (red) and alkaline butt (red) or no change (orange/red) butt
Triple sugar iron test. No carbohydrate fermentation has occurred. Peptones are catabolized resulting in an alkaline pH due to the production of ammonia.
Medium has sodium thiosulfate and Ferris sulfate to produce blackening.
Triple sugar iron test. hydrogen sulfide production.
Positive result produces a cherry red reagent layer.
Indole production test
Glucose fermentation reaction with methyl red PH reagent. Positive test is red. Negative test is yellow.
Methyl red test
Glucose fermentation. Red layer is a positive test.
Voges-proskauer test
Positive tube shows growth on slanted surface
Citrate utilization test
Prevents transpeptideation resulting in a weakened peptidoglycan
Penicillin
Has an affinity for bacterial ribosomes causing missreading of codons in mRNA interfering with protein synthesis.
Streptomycin
Has an affinity for bacterial ribosomes preventing peptide bond formation between amino acids during protein synthesis.
Chloramphenicol
Have an affinity for bacterial ribosomes prevent hydrogen bonding between the anti-codon on the transfer RNA amino acid complex and the codon on messenger RNA during protein synthesis.
Tetracyclines
Inhibits cell wall synthesis
Bactracin
Distraction of cell membrane
Polymyxin
Inhibits RNA synthesis
Rifampin
Inhibits DNA synthesis
Quinolone
