Team:BCCS-Bristol/Calendar-Notebook/21 July 2008

From 2008.igem.org

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Growth curve measurement

The OD₆₀₀ of E. coli MC1000 and MG1655 was measured over a day so that we would know whether the bacteria were in their log phase in any experiments requiring innoculation of bacteria.

A, B and C show 100, 500 and 200 µl innoculations of overnight culture in 50 ml LB broth, respectively.

Time (minutes)				OD₆₀₀ (1:5 diluted)
		MG1655				MC1000
	A	B	C		A	B	C
0	0.0016	0.0042	0.017		0.0016	0.0044	0.0174
60	0.008	0.013	0.073		0.006	0.017	0.068
80	0.011	0.052	0.109		0.009	0.046	0.123
100	0.019	0.059	0.142		0.017	0.067	0.167
120	0.029	0.076	0.169		0.024	0.083	0.183
140	0.039	0.095	0.213		0.039	0.144	0.226
160	0.063	0.136	0.26		0.053	0.147	0.275
180	0.074	0.166	0.276		0.069	0.186	0.298
200	0.105	0.207	0.301		0.089	0.22	0.316
220	0.134	0.25	0.355		0.116	0.268	0.331
240	0.178	0.276	0.386		0.151	0.293	0.341
260	0.233	0.329	0.419		0.198	0.325	0.372
280	0.239	0.334	0.435		0.203	0.326	0.375
300	0.263	0.363	0.473		0.238	0.34	0.388
320	0.296	0.391	0.491		0.27	0.364	0.412
340	0.319	0.425	0.511		0.29	0.378	0.429
360	0.32	0.435	0.512		0.294	0.382	0.432
380	0.392	0.542	0.514		0.311	0.434	0.438
400	0.435	0.549	0.588		0.348	0.457	0.497
420	0.47	0.52	0.487		0.382	0.462	0.504
440	0.483	0.586	0.633		0.477	0.437	0.516
460	0.498	0.482	0.629		0.411	0.484	0.517
480	0.574	0.643	0.721		0.451	0.516	0.524
500	0.607	0.66	0.73		0.455	0.514	0.515
520	0.65	0.673	0.723		0.453	0.522	0.523

BCCS-080721-growth curve E. coli MC1000 and MG1655.PNG

Swimming agar assay

Chemotaxis experiment for MG1655 and MC1000 showed no real difference between the control (water) and the aspartate. This may have been due to higher agar concentration caused by an unusually large water loss in autoclaving, causing high concentrations of agar. The experiment repetition was started today.

BioBrick Transformation

The transformation of bacteria with BioBrick from paper was started. Therefore, a hole was punched out of the DNA spot of a GFP generator and put in TE buffer (below) following protocol on iGEM sheet.

Miscellaneous

DMSO stocks of E. coli MC1000 and MG1655 were made as back up in the -80^oC freezer.