Team:Heidelberg/Notebook measure/NotebookFC

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Notebook

Contents

8-28-2009

  • First Flow cytometry measurement with cells transfected with cmv-GFP, jet-GFP and jet-cmv-GFP using jet and cmv as reference

9-9-2009

  • Second FC measurement with U2OS, MCF7 and HeLa with cmv-GFP, jet-GFP and jet-cmv-GFP using jet and cmv as reference
  • First measurement of inducible promoters in HeLa

9-11-2009

  • Third FC measurement with U2OS, MCF7 and HeLa with cmv-GFP, jet-GFP and jet-cmv-GFP using only jet as reference
File:HD09 Hannah FACSHeLa110909.JPG
Figure 1: Flow cytometry results of MCF7 GFP fluorescence intensity normalized with mCherry transfected in a 1:2 ratio.
File:HD09 Hannah HeLa110909.JPG
Figure 2: Flow cytometry results of MCF7 GFP fluorescence intensity normalized with mCherry transfected in a 1:2 ratio.
File:HD09 Hannah FACSU2OS110909.JPG
Figure 3: Flow cytometry results of U2OS GFP fluorescence intensity normalized with mCherry transfected in a 1:2 ratio.
File:HD09 Hannah U2OS110909.JPG
Figure 4: Flow cytometry results of U2OS GFP fluorescence intensity normalized with mCherry transfected in a 1:2 ratio.

9-21-2009

  • We decided to change our GFP:Cherry ratio from 1:2 to 2:1 to ensure that we have GFP in every cell expressing mCherry and thereby increasing the number of mCherry cells that also contain GFP.
  • Time course measurement of 24 and 96 well plate. Triplicates of the same transfection reaction were loaded at different positions in the plate to check wether the results change during the time course of the experiment.
Figure 1: Flow cytometry results of HeLa GFP fluorescence intensity normalized with mCherry transfected in a 2:1 ratio.
File:HD09 Hannah 24wellTime.JPG
Figure 2: Time measurement 24 Well Flow cytometry results of HeLa GFP fluorescence intensity normalized with mCherry transfected in a 1:2 ratio.
Figure 3: Flow cytometry results of HeLa GFP fluorescence intensity normalized with mCherry transfected in a 1:2 ratio.
File:HD09 Hannah 96wellTime.JPG
Figure 4: Time measurement 96 Well Flow cytometry results of HeLa GFP fluorescence intensity normalized with mCherry transfected in a 1:2 ratio.

9-24-2009

  • We performed standard measurements in HeLa, U2OS and MCF7 with our three standard promoters: CMV, JeT and JeT-CMV. These measurements will be performed in 3 independent measurements. The initial result of U2OS is shown below, which includes the following samples (see table 1).


Sample
negative control (transfection with a non-fluorescent plasmid p6)
cotransfection with JeT plasmid and p6
cotransfection with JeT_CMV plasmid and p6
cotransfection with CMV plasmid and p6
cotransfection with mCherry reference plasmid and p6
cotransfection with CMV plasmid and mCherry reference plasmid
cotransfection with JeT plasmid and mCherry reference plasmid
cotransfection with JeT_CMV plasmid and mCherry reference plasmid
Figure ?: Flow cytometry results of the U2-OS standard plate The .
  • We measured the constitutive promoters in the HeLa cell line. Unfortunately, the number of the viable cells are not enough for a reliable measurement.


9-25-2009

  • We measured the MCF-7 Standard plate, which includes the same samples as above (see table 1)
Figure ?: Flow cytometry results of the MCF-7 standard plate The .


9-28-2009

  • We measured the MCF-7 and the U2-OS Standard plate again (same samples).
Figure ?: Flow cytometry results of the U2-OS standard plate The .
Figure ?: Flow cytometry results of the MCF-7 standard plate The .

9-29-2009

  • We measured the MCF-7 and the U2-OS Standard plate the third time (same samples).
Figure ?: Flow cytometry results of the U2-OS standard plate The .
Figure ?: Flow cytometry results of the MCF-7 standard plate The .

9-30-2009

  • We measured the constitutive promoters in the HeLa cell line again.

10-02-2009

  • We measured the U2-OS NFkB plate 10 hours after induction.

10-03-2009

  • We measured the HIF promoter in MCF-7. Unfortunately, the cell number was too low.

10-05-2009

  • We measured the HeLa Standard plate in three different experiments????, which includes the same samples as above (see table 1)
  • We measured the constitutive promoters in the HeLa cell line again. Unfortunately, the number of the viable cells are not enough for a reliable measurement.
  • We measured the AhR ??? promoter in HeLa. Unfortunately, the number of the viable cells are not enough for a reliable measurement.

10-07-2009

  • We measured the U2-OS NFkB plate 10 hours after induction again.

10-08-2009

  • We measured the Estrogen promoter in MCF-7. Unfortunately, the number of the viable cells are not enough for a reliable measurement.

10-09-2009

  • We measured the HeLa Standard plate (fourth time).
  • We measured the SREBP ??? in HeLa.
  • We measured the constitutive promoters in HeLa again.

10-12-2009

  • We measured the HeLa Standard plate (fifth time).
  • We measured the promosing NKkB promoter NIIL10 ??? in different medium in U2-OS. Here, we want to see, if there is an induction of a nonspecific drug (Thiazolidin??) and how the different medium affect the promoter activity. No significant change.

10-13-2009

  • We measured the p53 ?? promoter in MCF-7. Unfortunately, the flow cytometry machine broke up the measurement and so we didn't have all the important data.

10-14-2009

  • We measured


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