From 2007.igem.org

09/10/07

• Miniprep for

-[http://partsregistry.org/Part:BBa_I763027 I763027];

-[http://partsregistry.org/Part:BBa_I763028 I763028];

-[http://partsregistry.org/Part:BBa_I763019 I763019];

-[http://partsregistry.org/Part:BBa_I763021 I763021]+[http://partsregistry.org/Part:BBa_P0412 P0412];

-[http://partsregistry.org/Part:BBa_I763021 I763021]+[http://partsregistry.org/Part:BBa_S03520 S03520];

-[http://partsregistry.org/Part:BBa_I763021 I763021]+[http://partsregistry.org/Part:BBa_S0100 S0100];

Digestion for:

-[http://partsregistry.org/Part:BBa_I763027 I763027] with Xba/Spe;

-[http://partsregistry.org/Part:BBa_I763028 I763028] with Spe/Pst1 and with Xba/Spe;

-[http://partsregistry.org/Part:BBa_I763019 I763019] with Xba/Spe;

-[http://partsregistry.org/Part:BBa_I763021 I763021]+[http://partsregistry.org/Part:BBa_P0412 P0412] with Eco/Spe;

-[http://partsregistry.org/Part:BBa_I763021 I763021]+[http://partsregistry.org/Part:BBa_S03520 S03520] with Eco/Spe;

-[http://partsregistry.org/Part:BBa_I763021 I763021]+[http://partsregistry.org/Part:BBa_S0100 S0100] with Eco/Spe;

• Band extraction from gel for all digestion;
• We have problems with [http://partsregistry.org/Part:BBa_J52034 J52034] and with [http://partsregistry.org/Part:BBa_I763019 I763019].
• Ligations for:

-[http://partsregistry.org/Part:BBa_I763028 I763028] + [http://partsregistry.org/Part:BBa_I763007 I763007]

-[http://partsregistry.org/Part:BBa_S0100 S0100] + [http://partsregistry.org/Part:BBa_J04431 J04431], because we want to understand if LacI operates well;

-[http://partsregistry.org/Part:BBa_J06550 J06550] + [http://partsregistry.org/Part:BBa_J04631 J04631], with this ligation we want to understand if it operates well, because it doesn't leak.

09/11/07

• Digestion for:

-[http://partsregistry.org/Part:BBa_I763020 I763020] with Xba/Pst1;

-[http://partsregistry.org/Part:BBa_J22101 J22101] with Xba/Pst1;

• Testing our devices

We take 5 ml of LB culture ground in which we put a colony of bacteria which contain Ptet-LacI Plac-cI-LacY-GFP plasmid.

We examine a drop of it using our fluorescence microscopy setup. In these conditions, the bacteria don’t beam fluorescence.

Starting from this moment, we put into the culture ground 1 mM of IPTG every 30 minutes until it is 4 millimolar, checking each time any eventual fluorescence, with negative results.

After diluting 1 ml of the previous one with 4 ml of LB, we add IPTG until 1 ml of our new solution is 10 mM, then 20 mM and finally 40 mM. Despite that, no fluorescence is visible.

Every time we checked for fluorescence, we used not only the photo camera but also the photomultiplier; every measurement showed a 0.16-0.18 volts offset, despite the bacteria being non-fluorescent: at least, we regulated that offset!

09/12/07

09/13/07

09/14/07

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