- We check growth curve for bacteria containing an intermediate plasmid.
- Transformation for C0012, J52034 and B0015.
- Ligation R0051+I13507 (I763007) (vector 2ul, insert 6ul) and ligation J04500+ C0051 (I763005)(vector 2ul, insert 10ul).
- There are no colonies for C0012, thus we inoculate I52034 and B0015 colonies in 5ml.
- We strake on plates 30/07 ligations.
- We go along with the 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 8th August.
-B0015 digestion with Xba/Pst1;
-I52034 digestion with Spe/Pst1;
- Gel extraction (we can’t see B0015);
-Control digestion of 5 ul with Eco/Pst1;
-A new B0015 digestion with Xba/Pst1;
-Run on electrophoresis gel. We have problems with gel because we don’t see the band;
- J22101 digestion with Xba/Pst1 ;
-Glycerol stocks preparation.