Team:Freiburg/Notebook/10 September

From 2011.igem.org

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===ligation for cloning PcpcG infront of CFP/YFP===
===ligation for cloning PcpcG infront of CFP/YFP===
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Ligation
 +
 +
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1. add H2O 11 μl
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 +
2. add 2 μl Ligase Buffer 10x
 +
 +
3. add Insert 1
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 +
4. add Vector (20ng needed. When proceeding from 3A digestion use 2 μl)
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 +
5. Add 1 μl T4-DNA Ligase
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6. Incubate 30 min at room temperature
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7. heat for 20 minutes at 80°C
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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;"|  
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| style="border:0.0139in solid #808080;padding:0.0194in;"| Name of part
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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;"| 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;"|  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;"|  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;"|  11 μl
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|}
==<span style="color:blue;">blue light receptor</span>==
==<span style="color:blue;">blue light receptor</span>==

Revision as of 15:55, 11 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


  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)

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


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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