Week 12
From 2007.igem.org
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'''Testing our devices''' | '''Testing our devices''' | ||
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| + | We want to test if our newly-constructed devices work, that is if they beam flourescence when inducted with IPTG. | ||
*In 2 different tubes we add 5ml of LB medium and 2 different colonies of [http://partsregistry.org/Part:BBa_I763019 I763019] plasmid. | *In 2 different tubes we add 5ml of LB medium and 2 different colonies of [http://partsregistry.org/Part:BBa_I763019 I763019] plasmid. | ||
*In another tube we add 5ml of LB medium and a colony of PLac-cI-GFP plasmid. | *In another tube we add 5ml of LB medium and a colony of PLac-cI-GFP plasmid. | ||
*After 2 hours we measure the OD value and it is around 0.5. | *After 2 hours we measure the OD value and it is around 0.5. | ||
| - | * For each original tube we divide the bacteria fluid in two different tubes. | + | *For each original tube we divide the bacteria fluid in two different tubes. |
| - | * We add | + | *We add 1mM IPTG to the solution into one tube. |
*The analyzed fluid without IPTG includes: | *The analyzed fluid without IPTG includes: | ||
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-2.5ul of kanamicin, | -2.5ul of kanamicin, | ||
| - | -50ul of IPTG. | + | -50ul of 100mM IPTG. |
| - | *We examine a bacteria fluid drop from of | + | *We examine a bacteria fluid drop from of each tube with our fluorescence microscope. |
*The bacteria with [http://partsregistry.org/Part:BBa_I763019 I763019] plasmid don’t beam fluorescence with and without IPTG; | *The bacteria with [http://partsregistry.org/Part:BBa_I763019 I763019] plasmid don’t beam fluorescence with and without IPTG; | ||
| - | *Very few bacteria with PLac-cI-GFP plasmid and without | + | *Very few bacteria with PLac-cI-GFP plasmid with and without IPTG beam fluorescence. |
| - | + | ||
| - | + | In conclusion, we see that bacteria with [http://partsregistry.org/Part:BBa_I763019 I763019] plasmid don't seem to work at all, and bacteria with PLac-cI-GFP plasmid don't seem to work properly. | |
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Revision as of 09:06, 3 October 2007
- 09/17/07
- Ligations for:
- We transform ligations and strake them on plates.
- 09/18/07
- We inoculate a colony for yesterday ligations in 5ml of LB medium O/N.
- 09/19/07
- Miniprep for:
-I763028;
-Ptet-LacI-T-PLac-GFP;
-Ptet-LacI-GFP(Spe/Pst1), (Xba/Pst1).
- Digestion for:
-I763028 with Eco/Spe;
-Ptet-LacI-T-PLac-GFP with Eco/Spe;
-Ptet-LacI-GFP with Eco/Spe;
-I763007 with Eco/Xba;
-Plac-cI-LacY with Eco/Spe1;
- Band extraction from gel for all digestion and then we observe:
-I763028, Ptet-LacI-T-PLac-GFP are died;
-Ptet-LacI-GFP is correct;
-I763007 not found;
-Plac-CI-LacY is correct.
- 09/20/07
Testing our devices
We want to test if our newly-constructed devices work, that is if they beam flourescence when inducted with IPTG.
- In 2 different tubes we add 5ml of LB medium and 2 different colonies of I763019 plasmid.
- In another tube we add 5ml of LB medium and a colony of PLac-cI-GFP plasmid.
- After 2 hours we measure the OD value and it is around 0.5.
- For each original tube we divide the bacteria fluid in two different tubes.
- We add 1mM IPTG to the solution into one tube.
- The analyzed fluid without IPTG includes:
-2.5ml of original tube fluid,
-2.5ml of LB medium,
-2.5ul of kanamicin.
- The analyzed fluid with IPTG includes:
-2.5ml of original tube fluid,
-2.5ml of LB medium,
-2.5ul of kanamicin,
-50ul of 100mM IPTG.
- We examine a bacteria fluid drop from of each tube with our fluorescence microscope.
- The bacteria with I763019 plasmid don’t beam fluorescence with and without IPTG;
- Very few bacteria with PLac-cI-GFP plasmid with and without IPTG beam fluorescence.
In conclusion, we see that bacteria with I763019 plasmid don't seem to work at all, and bacteria with PLac-cI-GFP plasmid don't seem to work properly.
- 09/21/07
