TUDelft/25 September 2008

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[http://2008.igem.org/TUDelft/13_October_2008 13] [http://2008.igem.org/TUDelft/14_October_2008 14] [http://2008.igem.org/TUDelft/15_October_2008 15] [http://2008.igem.org/TUDelft/16_October_2008 16] [http://2008.igem.org/TUDelft/17_October_2008 17] [http://2008.igem.org/wiki/index.php?title=TUDelft/18_October_2008&action=edit 18] [http://2008.igem.org/wiki/index.php?title=TUDelft/19_October_2008&action=edit 19]
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[http://2008.igem.org/wiki/index.php?title=TUDelft/27_October_2008&action=edit 27] [http://2008.igem.org/wiki/index.php?title=TUDelft/28_October_2008&action=edit 28] [http://2008.igem.org/wiki/index.php?title=TUDelft/29_October_2008&action=edit 29] [http://2008.igem.org/wiki/index.php?title=TUDelft/30_October_2008&action=edit 30] [http://2008.igem.org/wiki/index.php?title=TUDelft/31_October_2008&action=edit 31]

Contents

September 25th 2008=

Luciferase Assay

Sample preparation

All the cultures growing had reached an OD of about 0.4 in the morning. When 3 hours later the OD's had stayed practically constant, we lysed them. Also the cultures of K115036 growing at 30oC and 37oC have been lysed. Also has K115036 been pelleted and frozen at -20 for miniprepping later.

Of all cultures lysed, 10x and 100x dilutions have been made in H2O. 100x dilutions showed to be in the linear range of the BCA protein assay in a preliminary test, so these were used for total protein determinations.

Results

The strains measured contained one of the following constructs: P1010 in pSB1AT3, K115012, K115029, K115031, K115032 (which had a dubious PCR product, it was lane 32 1' here), K115034 and K115036. Only K115036 was not measured at 26oC.

Unfortunately the reader had some problems, making it impossible to do good measurements on K115031. Also did we find out pure 100x diluted lysis buffer gives quite high values in the total protein assay, so in next experiments we'll need to correct our calibration curve for this as well.

The normalized results of the luciferase assay (corrected for protein content) are displayed in graph 1.

TUDelft 080925Luciferase.jpg

Graph 1: Normalized Luciferase Activity measurements.