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Team:Evry/Notebook/Transposons/13.10.14
From 2014.igem.org
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== Obtaining the Kan gene on pNK2 == | == Obtaining the Kan gene on pNK2 == | ||
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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 first three well from new stock, next ones the old stock; oligo: F 82/ R 83; Tm tested: 55/60/65; elongation time: 1m30s |
<br> | <br> | ||
<img src="https://static.igem.org/mediawiki/2014/9/90/Evry_Kan_extract.JPG" width="50%"/> | <img src="https://static.igem.org/mediawiki/2014/9/90/Evry_Kan_extract.JPG" width="50%"/> | ||
Revision as of 21:46, 17 October 2014
== 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 first three well from new stock, next ones the old stock; 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.
