Team:Todai-Tokyo/Protocols/Notebook Sample

From 2009.igem.org

(Difference between revisions)
Line 34: Line 34:
<h2> Gel Purification </h2>
<h2> Gel Purification </h2>
<h2> RE Digest </h2>
<h2> RE Digest </h2>
 +
<h2> Colony PCR </h2>

Revision as of 02:44, 20 October 2009

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Contents

Miniprep

The following were miniprepped:

  • pLacI-RFP-dterm (From Ligation on 9/4; 4 colonies)
  • RFP ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J04450 BBa_J04450] from [http://partsregistry.org/assembly/libraries.cgi?id=19 2009 Spring Distribution]; 6 colonies)

PCR

The yqiT gene was PCR amplified from pLacI-yqiT-dterm (miniprepped on 9/7) using the following primers:

yqiT fwd: agcccgtgtagtactgtagagtt
yqiT rev: agggatgtttgccagtcgtaattag

using PCR program 1 and Ex-taq.

This PCR reaction attaches a Biobrick prefix and suffix to the gene.


Sequencing


Transformation

Ligation

Infusion

Gel Purification

RE Digest

Colony PCR

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