Team:Paris Saclay/Notebook/July/9

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For regulation system :
For regulation system :
*extraction of BioBrick BBa_K1155000 in concentrated medium for further DNA sequencing.
*extraction of BioBrick BBa_K1155000 in concentrated medium for further DNA sequencing.
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*the terminator BBa_B0010 in PSB1A2 plasmidwas extracted form iGEM plate kit 2013 and  was transformed into competent cells and then was cultured on solid medium
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*the plasmid PSB3K3(BBa_J04450) was extracted form iGEM plate kit 2013 and transformed into competent cells and then was cultured on solid medium for confirmation
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For sensor system:
For sensor system:
*A series of digestion, ligation were performed for BioBrick BphR2, BphR1, BphA1
*A series of digestion, ligation were performed for BioBrick BphR2, BphR1, BphA1
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Protocol : [[Team:Paris_Saclay/Protocols/Bacterial transformation|Bacterial transformation]]
Protocol : [[Team:Paris_Saclay/Protocols/Bacterial transformation|Bacterial transformation]]
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==='''B - PCB sensor system'''===
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===='''Objective : obtaining Bba_K1155001, Bba_K1155002 and BphR2 protein'''====
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===='''1 - Extraction of ...'''====
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...
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===='''1 - Digestion PCR products : '''====
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Zhou
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<br>
<br>

Revision as of 20:51, 22 September 2013

Navigation Home Team Project Labwork Notebook Human practices

Contents

Notebook : July 9

Summary :

For regulation system :

  • extraction of BioBrick BBa_K1155000 in concentrated medium for further DNA sequencing.


For sensor system:

  • A series of digestion, ligation were performed for BioBrick BphR2, BphR1, BphA1

Lab work

DNA purification

See protocol DNA purification Briobrick promoter BphR1, BphR2, BphA1 were purified.

Restriction digestion

We had 5 samples for digestion: 3 extracts, plasmid PSB1C3 and Plasmid PSB3K3(extracted from plate kit 2013 6F ).

For the plasmid (PSB1C3 and PSB3K3):

  • Plasmid : 4µl
  • (orange): 0.8µl
  • EcoR I: 0.5µl
  • PST I:0.5µl
  • H2O:2.2µl
  • Total:8µl

For the DNA extract:

  • DNA: 15µl
  • Buffer(orange): 3µl
  • EcoR I: 0.75µl
  • PST I:0.75µl
  • H2O:10.5µl
  • Total:30µl

After the digestion, superfluous enzyme was removed by Ethanol precipitation method. Then we suspended them with 10µl H2O.

Quantification

DNA concentration(ng/µl) 260/280
BphR2 24.2 1.97
BphA1 108.3 1.85
BphR1 97.9 1.82
PSB1C3 36.1 1.77
PSB3K3 19.4 1.77

Ligation

Common way for ligation.

  • Plasmid : 2µl
  • Bph : 2µl
  • Buffer : 2µl
  • T4 ligase : 12µl
  • H2O : about 7µl
  • Total : 20µl








Navigation Home Team Project Labwork Notebook Human practices

Notebook : July 9

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining Bba_K115507, Bba_K1155003

1 - Transformation of Bba_B0010

Abdou

Protocol : Bacterial transformation

Objective : obtaining biobricks in PSB3K3

1 - Transformation of Bba_J04450

Anaïs

Protocol : Bacterial transformation

B - PCB sensor system

Objective : obtaining Bba_K1155001, Bba_K1155002 and BphR2 protein

1 - Extraction of ...

...

1 - Digestion PCR products :

Zhou



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