噬菌体的来源、表现、鉴定、防治

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噬菌体的来源、表现、鉴定、防治：引用自（Marcin LOOE, Agata CZY, Eugenia SELL, Alicja WÊGRZYN, Peter NEUBAUER,
Grzegorz WÊGRZYN Appl. Genet. 45(1), 2004, pp. 111-120）英文
Common sources of phages in laboratories
– Dirty pipettes
– Dirty or wet cotton props
– Spoiled shaking incubators
– Strain and plasmid collections (test all incoming strains!)
– Filters of the sterile hood
– Room aeration systems with connections to different laboratories
– Electroporation cuvettes, if repeatedly applied and not carefully treated
– Electrode membranes and buffers that have been in contact with phage-infected cultures
Recognition of phage infection or prophage induction
– Lysis of cultures (unexpected decrease in optical density, low density after overnight
cultivation)
– Visible clumps and threads in the culture
– Thick white foam on the culture (even when the culture is dense), which does not disappear
if the flask is left for some time on the table
– Observation by light microscopy: clumps, fatty drops, dirty-like threads, no rod-shaped
cells
– Plaque test
– DNA chip technologies (including electric DNA chips)
Phage elimination methods
– Temperature treatment at 180°C for several hours under dry conditions
– Basic pH (10 M KOH, final pH of 10),
– Formaldehyde treatment or addition to the culture (especially for pipettes and other plastic
materials); increase in autoclaving time and/or temperature
– UV irradiation
– Attention: pore filter membranes are not able to exclude phages – use deep filters!
Preventive measures
– Sterile and clean working behaviour
– Dry boxes of pipette tips after autoclaving
– Treat all glass material covered by aluminium foil at 200°C overnight under dry conditions
– Use UV light regularly to disinfect the laboratory
– Use only clean pipettes (clean them regularly in formaldehyde)
– Use sterile cotton plugs for sterile cultivation (not aluminium foil to cover shake flasks);
do not use wet or dirty plugs
– Do not open flasks during the cultivation if not necessary. If you must open them, do it
only under sterile conditions
– Test your glycerol stocks for phages by the plaque test. Destroy spoiled cultures
– Autoclave old cultures! Treat them with strong disinfecting solutions
– Use basic washing solutions in your washing machine, as they inhibit phages
– Avoid moisture – dry all glass and plastic materials carefully
– Control regularly your sterile hood
– Clean your working tables regularly with disinfecting solutions and keep the laboratory
dust-free
Alleviation of effects of phage contamination
– Stop or inhibit bacterial growth as soon as you detect phage contamination in the culture
– Remove the carbon source from the medium

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