Team:Paris Saclay/Notebook/August/5

From 2013.igem.org

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* Gel : 1%
* Gel : 1%
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===='''Objective : obtaining Bba_K1155007'''====
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===='''1 - Digestion of Bba_I732017 by EcoRI/SpeI'''====
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Damir, Nadia
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Used quantities :
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* Bba_I732017 : 41µL
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* Buffer FD : 5µL
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* EcoRI : 2µL
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* SpeI : 2µL
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We let the digestion at 1h30 at 37°C.
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===='''2 -Electrophoresis to check the digestion of Bba_I732017 by EcoRI/SpeI'''====
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Damir, Nadia
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{|
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| style="width:350px;border:1px solid black;" |[[]]
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| style="width:350px;border:1px solid black;vertical-align:top;" |
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* Well 1 : 6µL of DNA Ladder
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* Well 2 et 3 : 5µL of Bba_I732017 digested by EcoRI/SpeI+1µL 6X loading dye
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* Gel : 0.8%
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|}
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Expected sizes :
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* RBS-LacZ : 3093 bp
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* PSB1A2 : 2079 bp
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{|
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| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
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We don't obtain fragments at the right size. We will do the digestion again.
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|}
===='''Objective : obtaining Bba_K1155004, Bba_K1155005, Bba_K1155006'''====
===='''Objective : obtaining Bba_K1155004, Bba_K1155005, Bba_K1155006'''====

Revision as of 17:04, 20 September 2013

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Contents

Notebook : August 5

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining Bba_K1155003

1 - Digestion of Bba_K1155003 by EcoRI/PstI

Nadia, XiaoJing

Used quantities :

  • Bba_K1155003 : 5µL
  • EcoRI FD : 1µL
  • PstI FD : 1µL
  • Buffer FD : 3µL
  • H2O : 20µL

We let the digestion at 37°C during 10 minutes ??????

2 - Electrophoresis of the digestion of Bba_K1155003

Damir

File:Ps.jpg|350px]]
  • Well 1 :
  • Well 2 : 6µL of DNA Ladder
  • Well 3 : 30µL of Bba_K1155003 from clone 9 digested by EcoRI/PstI+6µl of 6X loading dye
  • Well 4 : 5µL of Bba_K1155003 from clone 9+1µl of 6X loading dye
  • Well 5 : 30µL of Bba_K1155003 from clone 11 digested by EcoRI/PstI+6µl of 6X loading dye
  • Well 6 : 5µL of Bba_K1155003 from clone 11+1µl of 6X loading dye
  • Well 7 : 30µL of Bba_K1155003 from clone 12 digested by EcoRI/PstI+6µl of 6X loading dye
  • Well 8 : 5µL of Bba_K1155003 from clone 12+1µl of 6X loading dye
  • Gel : 1%

Objective : obtaining Bba_K1155007

1 - Digestion of Bba_I732017 by EcoRI/SpeI

Damir, Nadia

Used quantities :

  • Bba_I732017 : 41µL
  • Buffer FD : 5µL
  • EcoRI : 2µL
  • SpeI : 2µL

We let the digestion at 1h30 at 37°C.

2 -Electrophoresis to check the digestion of Bba_I732017 by EcoRI/SpeI

Damir, Nadia

[[]]
  • Well 1 : 6µL of DNA Ladder
  • Well 2 et 3 : 5µL of Bba_I732017 digested by EcoRI/SpeI+1µL 6X loading dye
  • Gel : 0.8%

Expected sizes :

  • RBS-LacZ : 3093 bp
  • PSB1A2 : 2079 bp

We don't obtain fragments at the right size. We will do the digestion again.

Objective : obtaining Bba_K1155004, Bba_K1155005, Bba_K1155006

1 - Colony PCR of Bba_K1155004, Bba_K115005, Bba_K1155006 in DH5α

Damir, Nadia, XiaoJing

COLONIES PIQUEES DANS 10µL d'eau par Tube !!!!!!!!!!!!!

Used quantities :

  • DNA : 2µL
  • Mix  : (it was divided in 25 tubes for each promotor with 23µL of mix in each on)
    • Oligo 44 : 3.5µL
    • Oligo 45 : 3.5µL
    • Buffer Dream Taq : 70µL
    • dNTP : 28µL
    • Dream Taq : 5µL
    • H2O : 590µL

PCR Program :

PsPCR0508.jpg


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