IGEM:IMPERIAL/2007/Projects/Experimental Design/Phase2: Difference between revisions
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<span class="_toggler_toggle-item1- | <span class="_toggler_toggle-item1-2-0"><span style="font-style:Bold; font-size:130%; text-indent: 5%;"> [+] 1.1 GFP Calibration Curve</span></span> | ||
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==1. | ==1.2 GFP Calibration Curve== | ||
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| align="left" width="170px" | | | align="left" width="170px" | | ||
| align="right" style="font-style:Bold; font-size:110%;" | [[IGEM:IMPERIAL/2007/Experimental Design/Phase2/Protocol 1. | | align="right" style="font-style:Bold; font-size:110%;" | [[IGEM:IMPERIAL/2007/Experimental Design/Phase2/Protocol 1.2 |'''Protocol''']] | ||
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| colspan=2 align="right" style="font-style:Bold; font-size:110%;" | [[IGEM:IMPERIAL/2007/Experimental Design/Phase2/Results 1. | | colspan=2 align="right" style="font-style:Bold; font-size:110%;" | [[IGEM:IMPERIAL/2007/Experimental Design/Phase2/Results 1.2 |'''Results''']] | ||
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'''Aims:''' <br> | '''Aims:''' <br> | ||
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<span class="_toggler_toggle-item1- | <span class="_toggler_toggle-item1-2-1">'''[+] Constant Conditions'''</span> | ||
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*Use a defined in vitro chassis, this will be determined after initial testing. It is most likely we will carry out a calibration curve 100ul commercial extract and a defined amount of home made cell extract | *Use a defined in vitro chassis, this will be determined after initial testing. It is most likely we will carry out a calibration curve 100ul commercial extract and a defined amount of home made cell extract | ||
*carry out at room temperature | *carry out at room temperature | ||
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<span class="_toggler_toggle-item1- | <span class="_toggler_toggle-item1-2-2">'''[+] Variables'''</span> | ||
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*[GFP] added to in vitro chassis varied | *[GFP] added to in vitro chassis varied | ||
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<span class="_toggler_toggle-item1- | <span class="_toggler_toggle-item1-2-3">'''[+] Sampling'''</span> | ||
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*Measure after addition of GFP to minimize degradation of GFP | *Measure after addition of GFP to minimize degradation of GFP | ||
Repetition: | Repetition: | ||
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<span class="_toggler_toggle-item1- | <span class="_toggler_toggle-item1-2-4">'''[+] Controls'''</span> | ||
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*Negative Control: In vitro systems only with no GFP added | *Negative Control: In vitro systems only with no GFP added | ||
*Positive Control: <font color=red> DEFINE </font> | *Positive Control: <font color=red> DEFINE </font> | ||
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<div style="float:right; font-size: 150%;"><span class="_toggler_show-toggle1- | <div style="float:right; font-size: 150%;"><span class="_toggler_show-toggle1-2 _toggler-toggler1-2a _toggler-toggler1-2b toggler1-2a">Show All Details</span><span class="_toggler_hide-toggle1-2 _toggler-toggler1-2a _toggler-toggler1-2b toggler1-2b" style="display:none;">Hide All Details</span></div> | ||
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<span class="_toggler_toggle-item1- | <span class="_toggler_toggle-item1-3-0"><span style="font-style:Bold; font-size:130%; text-indent: 5%"> [+] 1.3 Degradation Time of GFP</span></span> | ||
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==1. | ==1.3 Degradation Time of GFP== | ||
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<div style="position: relative; top: 5px;"> | <div style="position: relative; top: 5px;"> | ||
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{| border="0" align="right" style="font-style:Bold; font-size:140%; color:red;" | {| border="0" align="right" style="font-style:Bold; font-size:140%; color:red;" | ||
| align="left" width="170px" | | | align="left" width="170px" | | ||
| align="right" style="font-style:Bold; font-size:110%;" | [[IGEM:IMPERIAL/2007/Experimental Design/Phase2/Protocol 1. | | align="right" style="font-style:Bold; font-size:110%;" | [[IGEM:IMPERIAL/2007/Experimental Design/Phase2/Protocol 1.3 |'''Protocol''']] | ||
|- | |- | ||
| colspan=2 align="right" style="font-style:Bold; font-size:110%;" | [[IGEM:IMPERIAL/2007/Experimental Design/Phase1/Results 1. | | colspan=2 align="right" style="font-style:Bold; font-size:110%;" | [[IGEM:IMPERIAL/2007/Experimental Design/Phase1/Results 1.3 |'''Results''']] | ||
|}</div> | |}</div> | ||
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*To determine the half life of GFP for a range of temperatures | *To determine the half life of GFP for a range of temperatures | ||
<br> | <br> | ||
<span class="_toggler_toggle-item1- | <span class="_toggler_toggle-item1-3-1">'''[+] Constant Conditions:'''</span> | ||
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*50μl in vitro chassis | *50μl in vitro chassis | ||
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<span class="_toggler_toggle-item1- | <span class="_toggler_toggle-item1-3-2">'''[+] Variables:'''</span> | ||
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<br> | <br> | ||
*Temperature range: 4<sup>o</sup>C, 15<sup>o</sup>C, 25<sup>o</sup>C, 30<sup>o</sup>C, 37<sup>o</sup>C, 50<sup>o</sup>C | *Temperature range: 4<sup>o</sup>C, 15<sup>o</sup>C, 25<sup>o</sup>C, 30<sup>o</sup>C, 37<sup>o</sup>C, 50<sup>o</sup>C | ||
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*Degradation of GFP | *Degradation of GFP | ||
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<span class="_toggler_toggle-item1- | <span class="_toggler_toggle-item1-3-3">'''[+] Sampling:'''</span> | ||
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*Every 15 minutes. | *Every 15 minutes. | ||
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*3 repeats | *3 repeats | ||
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<span class="_toggler_toggle-item1- | <span class="_toggler_toggle-item1-3-4">'''[+] Controls:'''</span> | ||
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<br> | <br> | ||
*Negative Control: In vitro system only | *Negative Control: In vitro system only | ||
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<div style="float:right; font-size: 150%;"><span class="_toggler_show-toggle1- | <div style="float:right; font-size: 150%;"><span class="_toggler_show-toggle1-3 _toggler-toggler1-3a _toggler-toggler1-3b toggler1-3a">Show All Details</span><span class="_toggler_hide-toggle1-3 _toggler-toggler1-3a _toggler-toggler1-3b toggler1-3b" style="display:none;">Hide All Details</span></div> | ||
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Revision as of 11:57, 20 August 2007
1. Common GFP Experiments
[+] 1.1 Fluorescence dependence on volume
[+] 1.1 GFP Calibration Curve
[+] 1.3 Degradation Time of GFP
2. Project Specific Experiments
[+] 2.1 Cell by Date
[+] 2.2 Infector Detector