Team:Evry/Notebook/Transposons/13.10.14

From 2014.igem.org

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strategy:
strategy:
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Gel electrophoresis:
Gel electrophoresis:
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GEEEEEEEEEEEEEEEEEEEEEELLLLLLLLLL
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Purification sur gel:
Purification sur gel:
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PCR: enzyme: Q5; template: 150 ng pNK2; oligo: F 82/ R 83; Tm tested: 55/60/65; elongation time: 1m30s
PCR: enzyme: Q5; template: 150 ng pNK2; oligo: F 82/ R 83; Tm tested: 55/60/65; elongation time: 1m30s
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<img src="https://static.igem.org/mediawiki/2014/9/90/Evry_Kan_extract.JPG" width="50%"/>
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Purification sur gel:
Purification sur gel:

Revision as of 21:44, 17 October 2014

Picture

== BBa_1413044 retrieval of the transposon plasmid==
strategy:
Digestion of BBa_1413044 by BglII :
50 ul final volume, 5 ul Buffer NeB 3.1, 5 ul (20 ng/ul) BBa_1413044 in pSB1C3, 1 ul BglII, 39 ul H2O.
Gel electrophoresis:

Purification sur gel:
Kit thermoscientific "Gel extraction". Elution final in 30 ul.
== Obtaining the Kan gene on pNK2 ==
PCR: enzyme: Q5; template: 150 ng pNK2; oligo: F 82/ R 83; Tm tested: 55/60/65; elongation time: 1m30s

Purification sur gel:
Kit thermoscientific "Gel extraction". Elution final in 30 ul.

Oct 13