Team:Freiburg/Notebook/10 September

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<a href="https://2011.igem.org/Team:Freiburg/Notebook/9_September">Previous entry</a>
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<a href="https://2011.igem.org/Team:Freiburg/Notebook"> 10 September </a>
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==<span style="color:grey;">commons</span>==
 +
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===testdigest===
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'''Investigators:Julia'''
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 +
 +
[[File:Freiburg11 09 12tEST jULIA.Jpg| 400px|400px]]
==<span style="color:green;">green light receptor</span>==
==<span style="color:green;">green light receptor</span>==
-
===NAME OF YOUR EXPERIMENT===
+
===ligation for cloning PcpcG infront of CFP/YFP===
 +
Ligation
-
'''Investigators:NAME'''
 
 +
1. add H2O 11 μl
 +
2. add 2 μl Ligase Buffer 10x
-
==<span style="color:blue;">blue light receptor</span>==
+
3. add Insert 1
-
===NAME OF YOUR EXPERIMENT===
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4. add Vector (20ng needed. When proceeding from 3A digestion use 2 μl)
-
'''Investigators:NAME'''
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5. Add 1 μl T4-DNA Ligase
 +
6. Incubate 30 min at room temperature
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7. heat for 20 minutes at 80°C
-
==<span style="color:red;">red light receptor</span>==
 
-
===NAME OF YOUR EXPERIMENT===
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{| style="border-spacing:0;"
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| style="border-top:0.0139in solid #808080;border-bottom:0.0139in solid #808080;border-left:0.0139in solid #808080;border-right:none;padding:0.0194in;"| &nbsp;
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| style="border:0.0139in solid #808080;padding:0.0194in;"| Name of part
-
'''Investigators:NAME'''
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|-
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3. Transformation
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| style="border-top:none;border-bottom:0.0139in solid #808080;border-left:0.0139in solid #808080;border-right:none;padding:0.0194in;"| insert 1
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| style="border-top:none;border-bottom:0.0139in solid #808080;border-left:0.0139in solid #808080;border-right:0.0139in solid #808080;padding:0.0194in;"| &nbsp;4 μl PcpcG from PCR
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|-
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| style="border-top:none;border-bottom:0.0139in solid #808080;border-left:0.0139in solid #808080;border-right:none;padding:0.0194in;"| vector
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| style="border-top:none;border-bottom:0.0139in solid #808080;border-left:0.0139in solid #808080;border-right:0.0139in solid #808080;padding:0.0194in;"| &nbsp;2 μl [http://partsregistry.org/wiki/index.php?title=Part:BBa_I15016 BBa_I15016] /[http://partsregistry.org/wiki/index.php?title=Part:BBa_I15016 BBa_I1501]7
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|-
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| style="border-top:none;border-bottom:0.0139in solid #808080;border-left:0.0139in solid #808080;border-right:none;padding:0.0194in;"| H2O
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| style="border-top:none;border-bottom:0.0139in solid #808080;border-left:0.0139in solid #808080;border-right:0.0139in solid #808080;padding:0.0194in;"| &nbsp;11 μl
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|}
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==<span style="color:blue;">blue light receptor</span>==
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===Picking colonies===
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'''Investigators:Sophie'''
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inoculating cm-LB with clones from the Gibson-♥-NOT-Trafo.
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Labelled ♥-NOT-G a,b,c,d,e,f
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===Transformation===
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'''Investigators: Sophie'''
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 +
Transformation with the PR-♥-NOT-Ligation (6 different PR-vectors)
==<span style="color:orange;">Lysis cassette</span>==
==<span style="color:orange;">Lysis cassette</span>==
-
===NAME OF YOUR EXPERIMENT===
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===M48 in C3 Vector===
-
'''Investigators:NAME'''
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'''Investigators:Theo'''
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Part M48 was Minipreped and cut with EcoRI and PstI. pSB1C3 was also PCR amplified on Friday (this was probably the problem for the failed Transformation of the other part-project -see Notebook Sept 9th-) and cut with EcoRI and PstI, ligated with M48 and transformed into competent cells. The colonies were to be picked on Sunday.
 +
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<br>
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===Troubleshooting of lysis cassette===
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Colonies of the transformation done on Friday were picked and inoculated on Amp LB medium
==<span style="color:grey;">Precipitator</span>==
==<span style="color:grey;">Precipitator</span>==
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===NAME OF YOUR EXPERIMENT===
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===Inoculation with pbd-clones===
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'''Investigators: Sophie'''
-
'''Investigators: NAME'''
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I inoculated 200mL Cm-LB with clones containing the plastic binding domain. I want to extract the GFP-pbd protein to be able to make experiments with this protein and get data about the plastic binding performance
{{:Team:Freiburg/Templates/footer}}
{{:Team:Freiburg/Templates/footer}}

Latest revision as of 00:23, 22 September 2011


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

Contents

commons

testdigest

Investigators:Julia


Freiburg11 09 12tEST jULIA.Jpg

green light receptor

ligation for cloning PcpcG infront of CFP/YFP

Ligation


1. add H2O 11 μl

2. add 2 μl Ligase Buffer 10x

3. add Insert 1

4. add Vector (20ng needed. When proceeding from 3A digestion use 2 μl)

5. Add 1 μl T4-DNA Ligase

6. Incubate 30 min at room temperature

7. heat for 20 minutes at 80°C


3. Transformation
  Name of part
insert 1  4 μl PcpcG from PCR
vector  2 μl [http://partsregistry.org/wiki/index.php?title=Part:BBa_I15016 BBa_I15016] /[http://partsregistry.org/wiki/index.php?title=Part:BBa_I15016 BBa_I1501]7
H2O  11 μl

blue light receptor

Picking colonies

Investigators:Sophie

inoculating cm-LB with clones from the Gibson-♥-NOT-Trafo.

Labelled ♥-NOT-G a,b,c,d,e,f

Transformation

Investigators: Sophie

Transformation with the PR-♥-NOT-Ligation (6 different PR-vectors)

Lysis cassette

M48 in C3 Vector

Investigators:Theo


Part M48 was Minipreped and cut with EcoRI and PstI. pSB1C3 was also PCR amplified on Friday (this was probably the problem for the failed Transformation of the other part-project -see Notebook Sept 9th-) and cut with EcoRI and PstI, ligated with M48 and transformed into competent cells. The colonies were to be picked on Sunday.


Troubleshooting of lysis cassette

Colonies of the transformation done on Friday were picked and inoculated on Amp LB medium

Precipitator

Inoculation with pbd-clones

Investigators: Sophie

I inoculated 200mL Cm-LB with clones containing the plastic binding domain. I want to extract the GFP-pbd protein to be able to make experiments with this protein and get data about the plastic binding performance

                       

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