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An in vitro study into antibiotic combinations with enhanced activity against Escherichia coli

Rachel Hickman

Degree project in biology, Master of science (2 years), 2011 Examensarbete i biologi 30 hp till masterexamen, 2011

Biology Education Centre and Department of Cell and Molecular Biology, Uppsala University

Supervisor: Diarmaid Hughes

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coli cultures were grown in Mueller Hinton II (MHBII) broth (22g in a 1 litre of distilled water, autoclaved, pH 7.3 in accordance to Difco

TM

). The over-night cultures were used in the minimum inhibition concentration experiments, fractional inhibition concentration test experiments and the time-kill curve experiments. To create an overnight culture each were grown in 15mL falcon tubes with 2mL of MHBII with constant agitation to ensure good aeration at 37°C for 18 hours. The cultures in the minimum inhibition concentration tests and the fractional inhibition concentration tests were grown in 96-well microtiter plates without shaking at 37°C for 18 hours. Solid media was used in the time-kill curve experiments when samples were taken out and plated; the plates used were Muller Hinton agar plates (38g in a 1 litre of distilled water, autoclaved, pH 7.3 in accordance to Difco

TM

).

B*+9#9'+98,&Q,%74&

Ac&('.,$8!+&.%N!+.*6&).,!"YYY µg/mL, cefuroxime stock solution 2500 µg/mL, mecillinam stock solution 2000 µg/mL and phosphomycin stock solution 2000 µg/mL. Also LA plates containing 15µg/mL tetracycline were used for the storage of &($,+5.(8'C!9)*C!&7-'!!;)#*+')

<P"=RR!9)&3!WJJXI1D!TU#V!-*$+8)C. All antibiotics were obtained from Sigma-Aldrich.

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(20)

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$,.&3'(!$,&)4).&)%0!J+),:!$!'--',C.(5!86*&)%3$,,'*!-)-'&&'Q!"YY µL of MHBII was added to each well on the 96-well Microtiter plate, then "YY µL first antibiotic of "=!&)8'+!.5!&3'!

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References

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