Difference between revisions of "Team:TU Darmstadt/judging/interlab"

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<h5>Fluorescence and Absorbance 600 Measurement</h5>
 
<h5>Fluorescence and Absorbance 600 Measurement</h5>
 
<p>Fluorescence and absorbance was measured from six different devices, positive and negative control which are built in pSB1C3 containing chloramphenicol resistance. Samples were taken after 0,2,4,6 hours.<br>   
 
<p>Fluorescence and absorbance was measured from six different devices, positive and negative control which are built in pSB1C3 containing chloramphenicol resistance. Samples were taken after 0,2,4,6 hours.<br>   
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The arithmetic mean from two colonies was calculated and can be found below.  
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The arithmetic mean from two colonies was calculated and can be found below.
 
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<figure><center>
 
<figure><center>
 
<img src="https://static.igem.org/mediawiki/2017/b/bd/T--TU_Darmstadt--Interlab5.gif", alt="hier fehlt was", width=70%,>
 
<img src="https://static.igem.org/mediawiki/2017/b/bd/T--TU_Darmstadt--Interlab5.gif", alt="hier fehlt was", width=70%,>
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<figcaption> Fig. 4: Hier fehlt was!!!. </figcaption></center>
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<figcaption> Fig. 4: Fluorescence measurement of six different devices, positive and negative control.<br>
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Measurement settings for the plate reader: Excitation 485 nm, Emission 530 nm, 50 gains and 20 flashes. </figcaption></center>
 
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Revision as of 12:06, 26 October 2017

MainPage

InterLab Measurement Studies

iGEM is coping with the problem of different groups measuring fluorescence differently, in different units and evaluating the results non-identically. Therefore, the InterLab Study aims to develop a standardized protocol for repeatable fluorescence measurements of GFP. The study should be carried out and examined in different laboratories all over the world. Our team followed the standardized procedure guiding us through transformation, inoculation and measurement process.
For more information: Fourth International InterLab Measurement Study
Following below you can find our results.

Calibration

OD600 reference point

Several measurements of LUDOX-S40 and H²O were spectrometric analyzed below.

LUDOX-S40 H²O
Replicate 1 0,041700002 0,035300002
Replicate 2 0,042399999 0,035
Replicate 3 0,043000001 0,035700001
Replicate 4 0,043000001 0,034699999
Arithmetic Mean 0,042525001 0,035175
Corrected Absorbance 600 0,007350001
Reference OD600 0,0425
Correction Factor 5,78231249
Tab. 1: Absorbance measurement of LUDOX-S40 and H²O of four different replicates.


hier fehlt was
Fig. 1: Data from table 1 exported in beam chart.


Fluorescein standard curve:

The fluorescence of a dilution series, fluorescein in four replicates were measured in standard modes in our Tecan plate reader (infinite 200Pro).


hier fehlt was
Tab. 2: Fluorescence measurement of fluorescein in eleven dilutions and one control containing PBS.
Measurement settings for the plate reader: Excitation 485 nm, Emission 530 nm, 50 gains and 20 flashes.


Generated standard curve of fluorescence for different fluorescein concentrations can be found below.


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Fig. 2: Calibration curve of fluorescein: fluorescence measurement of different concentrations (µM).


hier fehlt was
Fig. 3: Calibration curve of fluorescein (log scale): fluorescence measurement of different concentrations (µM).

Cell Measurement

Fluorescence and Absorbance 600 Measurement

Fluorescence and absorbance was measured from six different devices, positive and negative control which are built in pSB1C3 containing chloramphenicol resistance. Samples were taken after 0,2,4,6 hours.
The arithmetic mean from two colonies was calculated and can be found below.


hier fehlt was
Fig. 4: Fluorescence measurement of six different devices, positive and negative control.
Measurement settings for the plate reader: Excitation 485 nm, Emission 530 nm, 50 gains and 20 flashes.


hier fehlt was
Fig. 5: Hier fehlt was!!!.


Fluorescein / OD600

Text!!!

hier fehlt was
Fig. 6: Hier fehlt was!!!.



Abschlusssatz!!!