Team:Freiburg/Notebook/16 July

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{{:Team:Freiburg/Templates/header}}
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==<span style="color:green;">green light receptor</span>==
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<html>
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<div id="notebook-page-header">
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<div id="notebook-back" width="100px" >
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<a href="https://2011.igem.org/Team:Freiburg/Notebook/15_July">Previous entry</a>
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</div>
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<div id="notebook-title">
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<a href="https://2011.igem.org/Team:Freiburg/Notebook"> 16 July </a>
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</div>
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<div id="notebook-next">
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<a href="https://2011.igem.org/Team:Freiburg/Notebook/18_July">Next entry</a>
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</div>
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</div>
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</html>
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===NAME OF YOUR EXPERIMENT===
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==<span style="color:red;">red light receptor</span>==
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'''Investigators:NAME'''
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===Miniprep===
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'''Investigators: Jakob'''
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==<span style="color:blue;">blue light receptor</span>==
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RZ Plasmid Miniprep
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===NAME OF YOUR EXPERIMENT===
 
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'''Investigators:NAME'''
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{| style="border-spacing:0;"
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name:
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Jakob
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Date:
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16.07.2011
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|-
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| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Continue from Experiment:
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==<span style="color:red;">red light receptor</span>==
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-14.07.2011
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-Ligation and transformation of ho1+terminator and PcyA+terminator
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|-
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| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Project Name:
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Red light receptor
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|}
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Procedure:
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The following procedures are carried out at a room temperature. All centrifugation steps should be performed between 11,000 – 16,000 x g
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1.Centrifuge 0.5 – 5 ml of bacterial culture in a clear 1.5 ml tube at full speed for 15 – 20 seconds in a microcentrifuge. Discard supernatant.
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2.Add 200 µl of '''P1 Buffer '''(Red) to the tube and resuspend pellet completely (i.e., by vortexing or pipetting).
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3.Add 200 µl of '''P2 Buffer''' (Green) and mix by inverting the tube 2 – 4 times. Cells are completely lysed when the solution appears clear, purple and viscous. '''Proceed to the next step within 1-2 minutes'''.
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4.Add 400 µl of '''P3 Buffer''' (Yellow) and mix gently but thoroughly. Do not vortex. The sample will turn yellow when the neutralization is complete. Allow the lysate to incubate at room temperature for 1-2 minutes.
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5. Centrifuge sample(s) for 2 minutes.
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6.Place a '''Zymo-Spin IIN''' column in a '''Collection Tube''' and transfer the supernatant from step 5into the '''Zymo-Spin IIN''' column. When pipetting the supernatant be careful not to disturb the green pellet to avoid transferring anz cellular debris to the column.
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7.Centrifuge the '''Zymo-Spin IIN/Collection Tube''' assembly for 30 seconds.
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8.Discard the flow-through in the '''Collection Tube, '''making sure the flow-through does not touch the bottom of the column. Return the '''Zymo-Spin IIN''' column to the '''Collection Tube'''
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9.Add 200 µl of '''Endo-Wash-Buffer''' to the column and centrifuge for 30 seconds.
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10. Add 400 µl of '''Plasmid Wash Buffer''' to the column. Centrifuge for 1 minute.
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11. Transfer the column into a clean 1.5 ml microcentrifuge tube and then add 30 µl of '''DNA Elution Buffer''' to the column. Centrifuge for 30 seconds to elute the plasmid DNA.
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===NAME OF YOUR EXPERIMENT===
 
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'''Investigators:NAME'''
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'''Documentation:'''
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* Parts: BBa_I15008 + B1006 and BBa_I1009 + B1006
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==<span style="color:orange;">Lysis cassette</span>==
 
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===NAME OF YOUR EXPERIMENT===
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* DNA concentration:
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'''Investigators:NAME'''
 
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{| style="border-spacing:0;"
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| DNA ng/
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 260/280
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 260/230
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|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| L22a(ho1+term)
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 90
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1,9
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1,99
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==<span style="color:grey;">Precipitator</span>==
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|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| L22b(ho1+term)
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 99,3
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1,85
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1,92
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===NAME OF YOUR EXPERIMENT===
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|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| L23a(PcyA+term)
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 45,6
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1,96
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2,42
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'''Investigators: NAME'''
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|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| L23b(PcyA+term)
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 58,7
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2,0
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2,58
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{{:Team:Freiburg/Templates/header}}
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|}
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<br/>
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* Labeled: l22a, l22b, l23a, l23b Blue rack -> freezer

Latest revision as of 00:57, 22 September 2011


This is the wiki page
of the Freiburger student
team competing for iGEM 2011.
Thank you for your interest!