Week 5

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Revision as of 13:28, 12 September 2007

07/30/07

• We check growth curve for bacteria containing an intermediate plasmid.
• Transformation for [http://partsregistry.org/Part:BBa_C0012 C0012], [http://partsregistry.org/Part:BBa_J52034 J52034] and [http://partsregistry.org/Part:BBa_B0015 B0015].
• Ligation [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)] (vector 2ul, insert 6ul) and ligation [http://partsregistry.org/Part:BBa_J04500 J04500]+ [http://partsregistry.org/Part:BBa_C0051 C0051] [http://partsregistry.org/Part:BBa_I763005 (I763005)](vector 2ul, insert 10ul).

07/31/07

• There are no colonies for [http://partsregistry.org/Part:BBa_C0012 C0012], thus we inoculate [http://partsregistry.org/Part:BBa_I52034 I52034] and [http://partsregistry.org/Part:BBa_B0015 B0015] colonies in 5ml.
• We strake on plates 30/07 ligations.
• We go along with the [http://partsregistry.org/Part:BBa_J04431 J04431] fluorescence test:

-From the glycerol stock we sprout 1ml at 37°C for 1h in agitation.

-Every 15 minutes we check bacterial OD and fluorescence. Cells are fluorescent at t= 210min and OD=0.5.

• We decide to repeat the test to understand if we can switch off the fluorescent cells with a higher amount of glucose.

We will perform this test on 07/08.

08/01/07

-Miniprep of [http://partsregistry.org/Part:BBa_I52034 I52034] and [http://partsregistry.org/Part:BBa_B0015 B0015];

-[http://partsregistry.org/Part:BBa_B0015 B0015] digestion with Xba/Pst1;

-[http://partsregistry.org/Part:BBa_I52034 I52034] digestion with Spe/Pst1;

- Gel extraction (we can’t see [http://partsregistry.org/Part:BBa_B0015 B0015]);

-[http://partsregistry.org/Part:BBa_R0015 R0015]+[http://partsregistry.org/Part:BBa_I13507 I13507][http://partsregistry.org/Part:BBa_I763007 (I763007)] ligation which doesn’t give colonies thus we transform with the other 10ul of ligation;

-[http://partsregistry.org/Part:BBa_J04500 J04500]+[http://partsregistry.org/Part:BBa_C0051 C0051][http://partsregistry.org/Part:BBa_I763005 (I763005)]ligation which gives colonies so we inoculate in 5ml;

-Transformation for [http://partsregistry.org/Part:BBa_C0012 C0012] (4ul) and for [http://partsregistry.org/Part:BBa_J22101 J22101] (2ul);

-A new [http://partsregistry.org/Part:BBa_R0051 R0051]+ [http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)] (vector 4ul, insert 8ul) ligation.

08/02/07

-Miniprep of [http://partsregistry.org/Part:BBa_J04500 J04500]+[http://partsregistry.org/Part:BBa_C0051 C0051] [http://partsregistry.org/Part:BBa_I763005 (I763005)];

-Control digestion of 5 ul with Eco/Pst1;

-A new [http://partsregistry.org/Part:BBa_B0015 B0015] digestion with Xba/Pst1;

-Run on electrophoresis gel. We have problems with gel because we don’t see the band;

-Band extraction for [http://partsregistry.org/Part:BBa_B0015 B0015] e [http://partsregistry.org/Part:BBa_I763005 I763005];

-There are no colonies for [http://partsregistry.org/Part:BBa_C0012 C0012], but there are colonies for [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507][http://partsregistry.org/Part:BBa_I763007 (I763007)]. So, we identify the correct ligation protocol;

- [http://partsregistry.org/Part:BBa_R0010 R0010], [http://partsregistry.org/Part:BBa_P0412 P0412], [http://partsregistry.org/Part:BBa_C0012 C0012] transformation;

-Inoculation in 5 ml for [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507] and for [http://partsregistry.org/Part:BBa_J22101 J22101];

- [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)] fluorescence test to check if the RFP filter works properly.

08/03/07

-Miniprep for [http://partsregistry.org/Part:BBa_J22101 J22101] and for [http://partsregistry.org/Part:BBa_R0051 R0051]+ [http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)];

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

- Control digestion of [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)];

- [http://partsregistry.org/Part:BBa_J04500 J04500]+[http://partsregistry.org/Part:BBa_C0051 C0051] [http://partsregistry.org/Part:BBa_I763005 (I763005)] and [http://partsregistry.org/Part:BBa_B0015 B0015]and [http://partsregistry.org/Part:BBa_J22101 J22101] and [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)] run on electrophoresis gel and [http://partsregistry.org/Part:BBa_J04500 J04500]+[http://partsregistry.org/Part:BBa_C0051 C0051] [http://partsregistry.org/Part:BBa_I763005 (I763005)] and [http://partsregistry.org/Part:BBa_J22101 J22101] extraction;

-Glycerol stocks preparation.

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