Template:Leanna newestuvlawn

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<table width=70%><tr><td><div id="bodybaby">the methods</div></td>
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<b>Prepare cell lawns for UV exposure.</b><br>
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&nbsp;&nbsp;&nbsp;&nbsp;<i>Escherichia coli</i> transformed with either pTSMa or pLPTa and a composite biobrick (one of K415006, K415013, K415022 or K415023) were grown 12-14 hours in 4mL aliquots of LB at 37&deg;C with 4uL of 1000x Kanamycin and Ampicillin and 35uL of 300uM IPTG until an OD measurement confirmed the cells to be in the log phase of growth. The remainder of the protocol was performed in the dark. The IPTG was washed out in two cycles of pelleting, decanting, and resuspending the pellet in the original volume of fresh LB. The cells were then pelleted and resuspended in 400uL of LB. Of this sample, 100uL were mixed with a 4mL aliquot of melted M9 minimal top agar () and kept at 50&deg;C while 4uL of 1000x Kan and Amp were added. If the composite biobrick was not K415022 or K415023, the mixture received 400uL of 300uM Acyl Homoserine Lactones (AHL). The mixture was vortexed, quickly poured into small petri dishes, and allowed to cool for 5 minutes. The plates then grew at 30&deg;C for 4 hours.
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<br><b>Expose cell lawns to UV.</b><br>
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Revision as of 06:47, 10 October 2010

MIT iGEM 2010

Methods

Bacterial lawn exposed to UV
Phage western blot
Mammalian Cell Lines
Microfluidic stress
bacterial lawn exposed to uv

Prepare cell lawns for UV exposure.
    Escherichia coli transformed with either pTSMa or pLPTa and a composite biobrick (one of K415006, K415013, K415022 or K415023) were grown 12-14 hours in 4mL aliquots of LB at 37°C with 4uL of 1000x Kanamycin and Ampicillin and 35uL of 300uM IPTG until an OD measurement confirmed the cells to be in the log phase of growth. The remainder of the protocol was performed in the dark. The IPTG was washed out in two cycles of pelleting, decanting, and resuspending the pellet in the original volume of fresh LB. The cells were then pelleted and resuspended in 400uL of LB. Of this sample, 100uL were mixed with a 4mL aliquot of melted M9 minimal top agar () and kept at 50°C while 4uL of 1000x Kan and Amp were added. If the composite biobrick was not K415022 or K415023, the mixture received 400uL of 300uM Acyl Homoserine Lactones (AHL). The mixture was vortexed, quickly poured into small petri dishes, and allowed to cool for 5 minutes. The plates then grew at 30°C for 4 hours.
Expose cell lawns to UV.