Week 12

From 2007.igem.org

(Difference between revisions)
Line 46: Line 46:
'''Testing our devices'''
'''Testing our devices'''
-
*In 2 different tubes we add 5ml of LB medium and 2 different colonies of 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. *After 2 hours we measure the OD value and it is around 0.5.  
+
*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.
* For each original tube we divide the bacteria fluid in two different tubes.
* We add the fluid without IPTG to one and the fluid with IPTG  to the other one.
* We add the fluid without IPTG to one and the fluid with IPTG  to the other one.
*The analyzed fluid without IPTG includes:
*The analyzed fluid without IPTG includes:
-
-2,5ml of original tube fluid,  
+
-2.5ml of original tube fluid,  
-
-2,5ml of LB medium,  
+
-2.5ml of LB medium,  
-
-2,5ul of kanamicin.
+
-2.5ul of kanamicin.
*The analyzed fluid with IPTG  includes:  
*The analyzed fluid with IPTG  includes:  
-
-2,5ml of original tube fluid,  
+
-2.5ml of original tube fluid,  
-
-2,5ml of LB medium,  
+
-2.5ml of LB medium,  
-
-2,5ul of kanamicin,
+
-2.5ul of kanamicin,
-50ul of IPTG.
-50ul of IPTG.
*We examine a bacteria fluid drop from of  each tube with our fluorescence microscopy.  
*We examine a bacteria fluid drop from of  each tube with our fluorescence microscopy.  
-
*The bacteria with 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 IPTG beam fluorescence;  
*very few bacteria with PLac-cI-GFP plasmid and without IPTG beam fluorescence;  
*several bacteria with PLac-cI-GFP plasmid with IPTG beam fluorescence.  
*several bacteria with PLac-cI-GFP plasmid with IPTG beam fluorescence.  
-
*After 3 hours we measured the OD value of each tubes and it is 1,4.  
+
*After 3 hours we measured the OD value of each tubes and it is 1.4.  
*Bacteria are too concentrate, thus we dilute them.  
*Bacteria are too concentrate, thus we dilute them.  
*We take 2ml of LB medium, 1ml of bacteria fluid with and without IPTG and 2ul of kanamicin.   
*We take 2ml of LB medium, 1ml of bacteria fluid with and without IPTG and 2ul of kanamicin.   

Revision as of 09:51, 21 September 2007

09/17/07
  • Ligations for:

-I763026 + I763019;

-I763026 + I763004;

-I763025 + J04031.

  • 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

  • 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 the fluid without IPTG to one and the fluid with IPTG to the other one.
  • 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 IPTG.

  • We examine a bacteria fluid drop from of each tube with our fluorescence microscopy.
  • The bacteria with I763019 plasmid don’t beam fluorescence with and without IPTG;
  • very few bacteria with PLac-cI-GFP plasmid and without IPTG beam fluorescence;
  • several bacteria with PLac-cI-GFP plasmid with IPTG beam fluorescence.
  • After 3 hours we measured the OD value of each tubes and it is 1.4.
  • Bacteria are too concentrate, thus we dilute them.
  • We take 2ml of LB medium, 1ml of bacteria fluid with and without IPTG and 2ul of kanamicin.
  • Examining with fluorescence microscopy and we saw that only bacteria with PLac-cI-GFP plasmid beame fluorescence.


09/21/07



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