Difference between revisions of "Template:IONIS Paris/Notebook/16-09-15"

 
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Liquid culture on 96 wells plate
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<i class="fa fa-angle-right"></i> When 0,6 < OD<sub>600</sub> < 0,8, induction with arabinose or IPTG
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<i class="fa fa-angle-right"></i> Incubate 5h at 30°C; for BBa_K325909, measure the kinetic during the 5h of induction and take the maximum value <br>
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<i class="fa fa-angle-right"></i> Add 50µL of NanoGlow substrate (Promega) for 50 uL of bacteria <br>
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<i class="fa fa-angle-right"></i> Measure kinetic over 120 min and take maximum value for culture with BBa_K1159001 and T7-nanoluc
  
 
 
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<!-- DATE de l'experience !ID aussi -->
 
<!-- DATE de l'experience !ID aussi -->
 
<span class="cd-date" id="16-09-15"><br>16 Sept. 15</span>
 
<span class="cd-date" id="16-09-15"><br>16 Sept. 15</span>

Latest revision as of 15:30, 18 September 2015

Miniprep

Liquid culture (pSB1C3-Holin/Endolysin x4 and pSB1C3-pDawn x2)
Liquid culture from the bacterial growth kinetic experiment (pDawn-Holin/Endolysin x2)

Digestion

Mix digestion preparation:
Tube Miniprep pDawn-H/E
H2O MQ 2 μL 3 μL
Buffer 2.1 1 μL 1 μL
DNA 5 μL 5 μL
EcoRI 1 μL 1 μL
PstI 1 μL -

37°C, 1h

Expected results

Expected results

Results

Results

There were no expected profile of digestion excepted for the first and the third wells with pDawn-Holin/Endolysin.

Digestion

Mix digestion preparation:
Tube Biobrick
H2O MQ 5 μL
Buffer 2.1 1 μL
DNA 3 μL
EcoRI 1 μL

37°C, 1h

Liquid culture of bacteria transformed with pSB1C3-nanoluc (x5)

Liquid culture on 96 wells plate

When 0,6 < OD600 < 0,8, induction with arabinose or IPTG Incubate 5h at 30°C; for BBa_K325909, measure the kinetic during the 5h of induction and take the maximum value
Add 50µL of NanoGlow substrate (Promega) for 50 uL of bacteria
Measure kinetic over 120 min and take maximum value for culture with BBa_K1159001 and T7-nanoluc


16 Sept. 15