Team:Freiburg/Notebook/7 September

From 2011.igem.org

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===PCR===
===PCR===
'''Investigators: Sophie''' <br/>
'''Investigators: Sophie''' <br/>
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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: Sophie
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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: 06.09.11
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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 (Date)
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(Name)
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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: Gibson of ♥ and NOT
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|}
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PCR-Mixture for one Reaction:
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For a 50 µl reaction use
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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;"| 32,5µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| H<sub>2</sub>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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|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 10µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 5x Phusion Buffer
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| of Primer
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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;"| 2.5µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Primer fw
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| ♥: P97
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NOT: P99
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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;"| 2.5µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Primer dw
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| ♥: P98
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NOT: P100
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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;"| 1µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| dNTPs
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| of Template DNA
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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;"| 1µl
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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-Template
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| NoT, M45 (BBa_Q0400)
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original DNA from Edinburgh (BBa_K322999)
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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;"| 0.5 µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Phusion (add in the end)
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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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|}
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Digestion with Dpn I and purification with PCR purification kit
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What program do you use?
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First 20 cycles touchdown 69°C -0.4/cycle
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next 10 cycles touchdown 72°C -0.2 per cycle
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PCR products were loaded onto a gel, but there were no bands execpt for not3.
==<span style="color:red;">red light receptor</span>==
==<span style="color:red;">red light receptor</span>==

Revision as of 14:01, 7 September 2011


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

Contents

Commons

Ligation

Investigators: Sandra

Ligation of PR1-PR6 with RFP (S1b and S2a). PR1-PR6 were digested with antarctica before start of ligation. Start of ligation at 12:40. Incubation at room temperature for at least 3 hours.

Trafo

Investigators: Sandra

Trafo of:

  • PR1+GFP
  • PR2+GFP
  • PR3+GFP
  • PR4+GFP
  • PR5+GFP
  • PR6+GFP
  • PR1+50b
  • PR2+50b

green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME


blue light receptor

no colonies on the plates

There were no colonies on the plates with ligated parts of the 2A-assembly.

Miniprep

Investigators: Sophie
yesterday two tubes with LB were inoculated with cells from our S35 glycerol stock(BBa_K322999). Today minipreps were performed.

PCR

Investigators: Sophie

Name: Sophie


Date: 06.09.11
Continue from Experiment (Date)

(Name)

Project Name: Gibson of ♥ and NOT

PCR-Mixture for one Reaction:

For a 50 µl reaction use


32,5µl H20 Name
10µl 5x Phusion Buffer of Primer
2.5µl Primer fw ♥: P97

NOT: P99

2.5µl Primer dw ♥: P98

NOT: P100

1µl dNTPs of Template DNA
1µl DNA-Template NoT, M45 (BBa_Q0400)

original DNA from Edinburgh (BBa_K322999)

0.5 µl Phusion (add in the end)

Digestion with Dpn I and purification with PCR purification kit What program do you use?

First 20 cycles touchdown 69°C -0.4/cycle

next 10 cycles touchdown 72°C -0.2 per cycle


PCR products were loaded onto a gel, but there were no bands execpt for not3.

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

NAME OF YOUR EXPERIMENT

Investigators: NAME

                       

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