Week 12

From 2007.igem.org

(Difference between revisions)
Line 44: Line 44:
::'''09/20/07'''
::'''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.

Revision as of 09:49, 21 September 2007

09/17/07
  • Ligations for:

-[http://partsregistry.org/Part:BBa_I763026 I763026] + [http://partsregistry.org/Part:BBa_I763019 I763019];

-[http://partsregistry.org/Part:BBa_I763026 I763026] + [http://partsregistry.org/Part:BBa_I763004 I763004];

-[http://partsregistry.org/Part:BBa_I763025 I763025] + [http://partsregistry.org/Part:BBa_J04031 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:

-[http://partsregistry.org/Part:BBa_I763028 I763028];

-Ptet-LacI-T-PLac-GFP;

-Ptet-LacI-GFP(Spe/Pst1), (Xba/Pst1).

  • Digestion for:

-[http://partsregistry.org/Part:BBa_I763028 I763028] with Eco/Spe;

-Ptet-LacI-T-PLac-GFP with Eco/Spe;

-Ptet-LacI-GFP with Eco/Spe;

-[http://partsregistry.org/Part:BBa_I763007 I763007] with Eco/Xba;

-Plac-cI-LacY with Eco/Spe1;

  • Band extraction from gel for all digestion and then we observe:

-[http://partsregistry.org/Part:BBa_I763028 I763028], Ptet-LacI-T-PLac-GFP are died;

-Ptet-LacI-GFP is correct;

-[http://partsregistry.org/Part:BBa_I763007 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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