Team:Freiburg/Notebook/3 August

From 2011.igem.org

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==<span style="color:orange;">Lysis cassette</span>==
==<span style="color:orange;">Lysis cassette</span>==
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===NAME OF YOUR EXPERIMENT===
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Digestion of Quickchange
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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: Theo
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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: 03.08.2011
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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 03.08.2011
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Ready Lysis Cassette (Part S49) + RBS
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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: Ready Lysis Cassette (Part S49) + RBS
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|}
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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;"| 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;"| Buffer, NEB4
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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;"|
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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;"| 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;"| BSA (10x)
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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;"|
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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;"| 2,5 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;"| Enzym 1
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| DpnI
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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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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 37,5 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;"| DNA
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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;"| Quickchange PCR
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|}
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50 μl total volume
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Incubate for about 1,5h at 37°C + Heat inactivation at 80°C for 20min.
==<span style="color:grey;">Precipitator</span>==
==<span style="color:grey;">Precipitator</span>==

Revision as of 17:27, 15 August 2011


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

Contents

green light receptor


Testdigest

Investigators:Jakob, Julia

Name:

Jakob

Date:

02.08.2011

Continue from Experiment 02.08.2011

Miniprep

Project Name: green light receptor, testdigest of CcaR with BamHI

For one reaction you need: For Mastermix: Number of samples+2extra


4μl H2O 20
1μl Buffer, NEB4 5
1μl BSA (10x) 5
0,5 μl Enzym 1 1 BamHI
0,5 μl Enzym 2 -
3 μl DNA 3

10 μl total volume


4μl H2O 20
1μl Buffer, NEB4 5
1μl BSA (10x) 5
0,5 μl Enzym 1 1 EcoRI
0,5 μl Enzym 2 1 PstI
3 μl DNA 3

10 μl total volume


Give 3 μl of DNA in an eppi and add 7μl of the mastermix.

Incubate for about 1h at 37°C.


Add 1 μl Loading dye buffer and load the gel.

Take a picture of the gel, print picture and label the lanes!


Freiburg 11 8 3 2011 7 09 46 PM.jpg200px


CcaR digested with BamH1 are negativ

  • To-do: quickchange (scroll down)


Quickchange


Investigator:Jakob

PCR


Name:

Jakob

Date:

03.08.2011

Continue from Experiment:
Project Name:

Quickchange CcaR

PCR-Mixture for one Reaction:

For a 50 µl reaction use


32,5µl H20
10µl 5x Phusion Buffer
2.5µl Primer fw Primer: P12
2.5µl Primer dw Primer: P13
1µl dNTPs
1µl DNA-Template DNA: CcaR
0.5 µl Phusion (add in the end)



results of transformation from 2nd Aug

Investigators:Julia
transformation worked!

blue light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



red light receptor

Colony picking

Investigator:Jakob Picked some colonies for grow over night

  • to-do: Miniprep

Lysis cassette

Digestion of Quickchange


Name: Theo


Date: 03.08.2011
Continue from Experiment 03.08.2011

Ready Lysis Cassette (Part S49) + RBS

Project Name: Ready Lysis Cassette (Part S49) + RBS
5μl Buffer, NEB4
5μl BSA (10x)
2,5 μl Enzym 1 DpnI
37,5 μl DNA Quickchange PCR

50 μl total volume


Incubate for about 1,5h at 37°C + Heat inactivation at 80°C for 20min.

Precipitator

NAME OF YOUR EXPERIMENT

Investigators: NAME

                       

Retrieved from "http://2011.igem.org/Team:Freiburg/Notebook/3_August"