Week 5
From 2007.igem.org
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::'''07/31/07''' | ::'''07/31/07''' | ||
*There isn’t any colony for C0012, so we inoculate I52034 and B0015 colony in 5 ml. | *There isn’t any colony for C0012, so we inoculate I52034 and B0015 colony in 5 ml. | ||
| - | *We | + | *We strake on plates 30/07 ligations. |
*We get along with the J04431 fluorescence test: | *We get along with the J04431 fluorescence test: | ||
-From the glycerol stock we sprout 1 ml at 37°C for 1 hour in agitation. | -From the glycerol stock we sprout 1 ml at 37°C for 1 hour in agitation. | ||
Revision as of 12:44, 27 August 2007
- 07/30/07
- We control bacterial grew (link).
- Transformation for C0012 and J52034 and B0015.
- We get along with the ligation R0051+I13507 (I763007) (vector 2 ul, insert 6 ul) and the ligation J04500+ C0051 (I763005) (vector 2 ul, insert 10 ul).
- 07/31/07
- There isn’t any colony for C0012, so we inoculate I52034 and B0015 colony in 5 ml.
- We strake on plates 30/07 ligations.
- We get along with the J04431 fluorescence test:
-From the glycerol stock we sprout 1 ml at 37°C for 1 hour in agitation.
-We verify the OD and the fluorescence every 15 minutes. The fluorescence compare at the istance t=210min and at the OD=0.5.
- At the end of this test we decide to repeat it to see if we can switch off the fluorescent cells with a major quantity of glucose to confirm our assumptions.
We will attend this test on 07/08.
- 08/01/07
- We get along with:
-Miniprep of I52034 and B0015;
-Digestion of B0015 with Xba/Pst1;
-Digestion of I52034 with Spe/Pst1;
-Extraction from gel (we can’t see B0015);
-R0015+I13507 (I763007) ligation which doesn’t give colonies so we transform with the other 10 ul of ligation;
-J04500+C0051 (I763005) ligation which gives colonies so we inoculate in 5ml;
-Transformation for C0012 (4ul) and for J22101 (2ul);
-A new R0051+ I13507 (I763007) (vector 4ul, insert 8ul) ligation.
- 08/02/07
- Today we do:
-Miniprep of J04500+C0051 (I763005);
-Control digestion of 5 ul with Eco/Pst1;
-A new B0015 digestion with Xba/Pst1;
-Digested run on electroforesi gel but we have problems with gel because we don’t see the band;
-Band exstraction for B0015 e I763005;
-We observe thre aren’t colonies for C0012, but there are colonies for R0051 + I13507 (I763007), so we identify the correct ligation protocol;
- R0010, P0412, C0012 transformation;
-Inoculation in 5 ml for R0051 + I13507 and for J22101;
- R0051 + I13507 (I763007) fluorescence test to verify if the filter for RFP is adapted, but it isn’t.
- 08/03/07
