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Team:Evry/Notebook/Transposons/13.10.14
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== BBa_1413044 retrieval of the transposon plasmid== | == BBa_1413044 retrieval of the transposon plasmid== | ||
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Digestion of BBa_1413044 by BglII : | 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. | + | <br>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. |
| - | + | <br> | |
Gel electrophoresis: | Gel electrophoresis: | ||
| - | + | <br> | |
GEEEEEEEEEEEEEEEEEEEEEELLLLLLLLLL | GEEEEEEEEEEEEEEEEEEEEEELLLLLLLLLL | ||
<img src="" width="50%"/> | <img src="" width="50%"/> | ||
| - | + | <br> | |
Purification sur gel: | Purification sur gel: | ||
| - | + | <br> | |
Kit thermoscientific "Gel extraction". Elution final in 30 ul. | Kit thermoscientific "Gel extraction". Elution final in 30 ul. | ||
| - | + | <br> | |
== Obtaining the Kan gene on pNK2 == | == Obtaining the Kan gene on pNK2 == | ||
| - | + | <br> | |
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 | ||
| - | + | <br> | |
<img src="" width="50%"/> | <img src="" width="50%"/> | ||
| - | + | <br> | |
Purification sur gel: | Purification sur gel: | ||
| - | + | <br> | |
Kit thermoscientific "Gel extraction". Elution final in 30 ul. | Kit thermoscientific "Gel extraction". Elution final in 30 ul. | ||
| - | + | <br> | |
<br>. | <br>. | ||
Revision as of 19:58, 17 October 2014
== BBa_1413044 retrieval of the transposon plasmid==
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:
GEEEEEEEEEEEEEEEEEEEEEELLLLLLLLLL
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.
.
