Team:Paris Saclay/Notebook/August/14

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*Well 1 : 6µL DNA Ladder
*Well 1 : 6µL DNA Ladder

Revision as of 22:00, 19 August 2013

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Contents

Notebook : August 14

Lab work

A - Aerobic/Anaerobic regulation system

Obtaining the NarK or NarG or NirB _RBS-LacZ-term in PSB1C3 and in PSB3K3

1 - Gel electrophoresis to check the digestion of Bba_K1155004,Bba_K1155005, Bba_K1155006 by EcoRI and SpeI

Anaïs, Nadia

IMAGE
  • Well 1 : 6µL DNA Ladder
  • Well 2 : 5µL of BBa_K1155006 digested by EcoRI/SpeI
  • Well 3 : 5µL of BBa_K1155005 digested by EcoRI/SpeI
  • Well 4 : 5µL of BBa_K1155004 digested by EcoRI/SpeI
  • Gel : 1%

Expected sizes :

  • PSB1C33 : 2070kb
  • NarK, NarG, NirB : 200kb 
We obtained fragments of the right size but in very few quantity. We do it again but this time we will use more quantity of enzymes AND ADN ?!
2 - Digestion of Bba_K1155004,Bba_K1155005, Bba_K1155006 by SpeI and EcoRI/SpeI

Anaïs, Nadia

Protocol : Digestion

  • SpeI :
    • Buffer : 2µL
    • SpeI : 2µL
    • ADN : 15µL
    • H20 : 1µL
  • EcoRI/SpeI :
    • Buffer : 3µL
    • SpeI : 2µL
    • EcoRI : 2µL
    • ADN : 20µL
    • H20 : 3µL
PSgel1 1408.jpg
  • Well 1 : 6µL DNA Ladder
  • Well 2 : 5µL of BBa_K1155006 digested by EcoRI/SpeI
  • Well 3 : 5µL of BBa_K1155005 digested by EcoRI/SpeI
  • Well 4 : 5µL of BBa_K1155004 digested by EcoRI/SpeI
  • Gel : 1%


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