Team:Kyoto/Secretion/Notebook

From 2012.igem.org

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37℃<br>
37℃<br>
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Revision as of 20:49, 24 September 2012

Contents

September 18

Gel extraction

写真お願いします。

  • LacP-torA-GFP-DT(EcoR1, Xba1) 36.9μg/mL 1.37(260/280) 0.82(260/230)

Ligation

J23107-tatABCD-pspA-DT(EcoR1, Spe1) 14.2μg/mLLacP-torA-GFP-DT(EcoR1, Xba1) 36.9μg/mLLigation High Ver.2total
81.59.519
LacP-torA-GFP-DT(EcoR1, Xba1) 36.9μg/mLLigation High Ver.2total
1.51.53

at 4℃ for 1 hour

Transformation

  • J23107-tatABCD-pspA-DT-LacP-torA-GFP-DT
  • LacP-torA-GFP-DT(EcoR1, Xba1)

Observation through a confocal microscope

We made sample for observation through a confocal microscope We incubated sample with 0mM/0.5mM IPTG for 1.5 hours. Then, we incubated onto medium without IPTG for 2.5 hours.

IPTG(mM)medium
0LB
0.1LB
0.1LB with 0.1mM IPTG
0LB with 2 percent glucose
0.1LB with 2 percent glucose
0LB with 5 percent glucose
0.1LB with 5 percent glucose
0LB with 2 percent glucose
0LB ここ途中
0LB
0LB

Then, we incubated onto medium without IPTG for 2.5 hours.




Restriction enzyme processing

J23101-Kil-DTBufferMXbaIPstBSAMilliQtotal
153113730
J23101-Kil-DTBufferMXbaIBSAMilliQtotal
310.514.510
J23101-Kil-DTBufferHPstMilliQtotal
310.55.510

37℃

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