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Team:Evry/Notebook/Transposons/15.10.14
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<h4> 14.10.2014 ligation </h4> | <h4> 14.10.2014 ligation </h4> | ||
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it gave only one clone. | it gave only one clone. | ||
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MB agar for <i>Pseudovibrio denitrificans</i> | MB agar for <i>Pseudovibrio denitrificans</i> | ||
<br>LB agar for LB for DH5alpha | <br>LB agar for LB for DH5alpha | ||
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<h4> Transformation <i>Pseudovibrio denitrificans</i> & Dh5 alpha with BBa_1413001 in BBa_1413044 </h4> | <h4> Transformation <i>Pseudovibrio denitrificans</i> & Dh5 alpha with BBa_1413001 in BBa_1413044 </h4> | ||
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Revision as of 22:44, 17 October 2014
14.10.2014 ligation
it gave only one clone.
Clone culture:
5 ml LB + 5 ul Kan (25 mg/ml) - 37 C°
Transformation Pseudovibrio denitrificans & Dh5 alpha with mRFP in BBa_1413044
electrocompetent cells:
1 mm cuvette; 1800 V
recuperation media: MB for Pseudovibrio denitrificans , LB for DH5alpha
plating:
MB agar for Pseudovibrio denitrificans
LB agar for LB for DH5alpha
Transformation Pseudovibrio denitrificans & Dh5 alpha with BBa_1413001 in BBa_1413044
electrocompetent cells:
1 mm cuvette; 1800 V
recuperation media: MB for Pseudovibrio denitrificans , LB for DH5alpha
plating:
MB agar for Pseudovibrio denitrificans
LB agar for LB for DH5alpha
Verification testing of pnK2 in Pseudovibrio denitrificans
16 clones of Pseudovibrio denitrificans were grown in 3 ml MB, 30 C°, 24h
DNA extraction:
Invitrogen DNA extraction kit used with the Gram - protocol.
nanodrops gave value between 80 ng/ul & 130 ng/ul
PCR for Kan gene insertion:
PCR: enzyme: Q5; template: 200 ng genomic preparation Pseudovibrio denitrificans; oligo: F 82/ R 83; Tm tested: 55; elongation time: 2m00s
PCR for Pseudovibrio denitrificans specific oligo:
PCR: enzyme: Q5; template: 200 ng genomic preparation Pseudovibrio denitrificans; oligo: F 72/ R 73; Tm tested: 55; elongation time: 2m00s
Oct 15
