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Team:Evry/Notebook/Transposons/13.10.14
From 2014.igem.org
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| - | + | <h4> BBa_1413044 retrieval of the transposon plasmid </h4> | |
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Kit thermoscientific "Gel extraction". Elution final in 30 ul. | Kit thermoscientific "Gel extraction". Elution final in 30 ul. | ||
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| - | + | <h4>Obtaining the Kan gene on pNK2</h4> | |
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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 | 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 | ||
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Kit thermoscientific "Gel extraction". Elution final in 30 ul. | Kit thermoscientific "Gel extraction". Elution final in 30 ul. | ||
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Revision as of 22:03, 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.
Oct 13
