Team:Freiburg/Notebook/Methoden

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(Difference between revisions)
(→Second Extension PCR)
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Pipette the following volumes into a PCR- tube:  
Pipette the following volumes into a PCR- tube:  
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</p><br>
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==<span style="color:#000000">PCR Purification==
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<p>
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bla
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</p><br>
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==<span style="color:#000000">Gel Run==
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<p>
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bla
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</p><br>
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==<span style="color:#000000">Ligation==
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<p>
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bla
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</p><br>
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==<span style="color:#000000">Colony PCR==
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<p>
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bla
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</p><br>
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==<span style="color:#000000">Mini-Prep==
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<p>
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bla
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</p><br>
</p><br>

Revision as of 17:18, 25 September 2012






First Extension PCR (Toolkit)

This step is necessary to add the the necessary restriction sites to the Direpeats. Pipette the following volumes into a PCR- tube:

Component Volume
Water, nuclease-free 15,75 µl
DMSO 0,75 µl
dNTPs 0,5 µl
Phusion Buffer 5,0 µl
Phusion Polymerase 0,25 µl
Water, nuclease-free up to 20 µl
Total vloume 20 µl



Second Extension PCR (Toolkit)

This step is necessary to add enzyme- binding –sites to the outer restriction sites. Use the product of the first extension- PCR as template. Pipette the following volumes into a PCR- tube:


PCR Purification

bla


Gel Run

bla


Ligation

bla


Colony PCR

bla




Mini-Prep

bla


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