Team:Freiburg/Notebook/30 August

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2.Ligation<br/>
2.Ligation<br/>
<br/>11µl H2O<br/>4µl PcpcG<br/>2µl pSB1C3<br/>2µl T4 Ligase buffer<br/> 1µl T4 Ligase<br/>
<br/>11µl H2O<br/>4µl PcpcG<br/>2µl pSB1C3<br/>2µl T4 Ligase buffer<br/> 1µl T4 Ligase<br/>
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Incubation for 3 hours at room temperature.<br/> heat inactivation at 80 degrees for 20 min.
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3. transformation
==<span style="color:blue;">blue light receptor</span>==
==<span style="color:blue;">blue light receptor</span>==

Revision as of 19:25, 30 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

Ligation of Promotor PcpcG in PSB1C3

Investigators:Julia

1. Digestion of PCR Product from yesterday[1]

38µl PCR product
5µl BSA (10x)
5µl NEB buffer 4
1µl EcoRI
1µlPstI

Incubation for 3 hours, afterwards the sample was cleaned with a PCR purification kit from Qiagen.

2.Ligation

11µl H2O
4µl PcpcG
2µl pSB1C3
2µl T4 Ligase buffer
1µl T4 Ligase

Incubation for 3 hours at room temperature.
heat inactivation at 80 degrees for 20 min.

3. transformation

blue light receptor

Transformation

...with ♥-A3-Not, ♥-A3-nOt and ♥-A3-noT </br>

Investigators: Sophie

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

Cloning

Investigators: Sophie
Project name: inducible promoter for pbd
again cloning of IPTG-inducible Promoter, new psb1C3-PCR product and ε5 (GFP-pbd) but this time I use Antarctic Phosphatase in the last hour of the digestion of the vector to avoid re-ligation. For 50μL reaction volume use 5,6μL Antarctic Phosphatase buffer and 1μL Antarctic Phosphatase.

                       

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