Team:University of Sheffield /Wet Lab

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(New page: ==Protocols== ===Tecan Fluorescence Measurement=== Protocol used for characterization : 1. Grow 5ml of overnight cultures of DH5-alpha cells containing the plasmid with GFP-LVA. 2. Re...)
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{| style="color:#888888;background-color:##888888;" cellpadding="5" cellspacing="2" border="2" bordercolor=#888888 width="85%" align="center"
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!align="center"|[[Team:University_of_Sheffield |Introduction]]
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!align="center"|[[Team:University_of_Sheffield /Project|Our project]]
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!align="center"|[[Team:University_of_Sheffield /Modelling|Modelling]]
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!align="center"|[[Team:University_of_Sheffield /Wet Lab|Wet Lab]]
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!align="center"|[[Team:University_of_Sheffield /Lab Books| Our team]]
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!align="center"|[[Team:University_of_Sheffield /Misc| Miscellaneous]]
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==Protocols==
==Protocols==

Revision as of 22:08, 27 October 2008

UniShefBanner.jpg


Introduction Our project Modelling Wet Lab Our team Miscellaneous


Protocols

Tecan Fluorescence Measurement

Protocol used for characterization :

1. Grow 5ml of overnight cultures of DH5-alpha cells containing the plasmid with GFP-LVA.

2. Resuspend the overnight cultures in 50 ml of LB medium until the OD600 reaches about 0.6.

3. Quickly add 0.2 mM of IPTG to the medium and plate that into 96 well plate.

4. To 96 well plate add 180 ul of LB + IPTG as a control, 180 ul of DH5-alpha cells not induced with IPTG, finally add 180 ul of DH5-alpha induced with IPTG. Carry out the aforementioned in duplicates.

5. Use the high flow cytometry machine Tecan to measure the fluorescence every 15 minutes for 8 hours (excitation wv – 485 nm, emission wv- 535 nm )(including shaking = aeration of the cultures 2 minutes prior each measurement)

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