IGEM:IMPERIAL/2007/Projects/Experimental Design/Phase2: Difference between revisions

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Test to determine the relationship between fluorescence and in vitro concentration of GFP. To test this purified samples of known [GFP] are added to in vitro chassis and the fluorescence measured. From this a calibration curve of [GFP] vs Fluorescence can be made. This can be used for data analysis to convert fluorescence into a [GFP].
Test to determine the relationship between fluorescence and in vitro concentration of GFP. To test this purified samples of known [GFP] are added to in vitro chassis and the fluorescence measured. From this a calibration curve of [GFP] vs Fluorescence can be made. This can be used for data analysis to convert fluorescence into a [GFP].
 
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'''Aims:'''
*To determine [GFP] vs Fluorescence 
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| colspan=2 align="right"  style="font-style:Bold; font-size:110%;" | [[IGEM:IMPERIAL/2007/Experimental Design/Phase2/Results 1.1 |'''Results''']]  
| colspan=2 align="right"  style="font-style:Bold; font-size:110%;" | [[IGEM:IMPERIAL/2007/Experimental Design/Phase2/Results 1.1 |'''Results''']]  
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'''Aims:'''
*To determine [GFP] vs Fluorescence 





Revision as of 12:49, 14 August 2007



1. Common GFP Experiments



1.1 GFP Calibration Curve


Status
Planned for 20/8/07 INCOMPLETE

Test to determine the relationship between fluorescence and in vitro concentration of GFP. To test this purified samples of known [GFP] are added to in vitro chassis and the fluorescence measured. From this a calibration curve of [GFP] vs Fluorescence can be made. This can be used for data analysis to convert fluorescence into a [GFP].

Protocol
Results

Aims:

  • To determine [GFP] vs Fluorescence





Constant Conditions:

(Show/Hide)


Variables:

(Show/Hide)


Sampling:

(Show/Hide)


Controls:

(Show/Hide)


Show All Details


1.2 Degradation Time of GFP


Status
Planned for 20/8/07 INCOMPLETE

Test the half life of GFP protein in an in vitro chassis. To test this a purified sample of known [GFP] are added to an in vitro chassis, then fluorescence will be measured at regular time intervals. The fluorescence will be converted into GFP molecules using the calibration curve. This will give; degradation of GFP as a function of time, from this the half life of GFP can be obtained. In addition, temperature may affect the half life of GFP and so the half life will be measured for an appropriate temperature range.

Protocol
Results

Aims:

  • To determine the half life of GFP for a range of temperatures

Conditions:

  • 50μl in vitro chassis
  • [GFP] added

Variables:

  • Temperature range: 4oC, 15oC, 25oC, 30oC, 37oC, 50oC
  • Range may change based upon the initial testing; will only test ranges that in vitro is stable at.
  • Degradation of GFP

Sampling:

  • Every 15 minutes.

Repetition:

  • 3 repeats

Controls:

  • Negative Control: In vitro system only
  • Positive Control: In vitro chassis with high concentration of purified GFP added at high temperature


2. Project Specific Experiments

[Cell by Date]

[Infector Detector]