Team:ESBS-Strasbourg/17 July 2008

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michi:
michi:
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*found paper PMID: 18385157 which mutates TATA box and quantifies transcription :-) (King of the day)
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*found paper PMID 18385157 which mutates TATA box and quantifies transcription :-) (King of the day)
=== Modeling ===
=== Modeling ===
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*The new BioBricks arrived in tubes with a sort of medium inside, we think they delivered us directly the bacteria with the BB-Plasmids inside, so we don't have to transform them on our own. Jippie!<br>
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*The new BioBricks arrived so we made an overnight culture of, in LB(Amp):
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**We made an overnight culture of, in LB(Amp):
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**p1010 in pSB1A3
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***p1010 in pSB1A3
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**BBa_c0012 (lacI)
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***BBa_c0012 (lacI)
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**BBa_c0051 (lamda cI)
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***BBa_c0051 (lamda cI)
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**BBa_j63001 (EYFP)
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***BBa_j63001 (EYFP)
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*We plated p1010 in pSB3K3 on LB Kan
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**We plated p1010 in pSB3K3 on LB Kan
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*Digestion of the plasmid of the BBa_J63002(?) clone with XbaI and SpeI
*Digestion of the plasmid of the BBa_J63002(?) clone with XbaI and SpeI

Latest revision as of 09:25, 20 July 2008

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Contents

DryLab

michi:

  • found paper PMID 18385157 which mutates TATA box and quantifies transcription :-) (King of the day)

Modeling

WetLab

  • Results Transformation
    • no clones for any of the three BioBricks
    • clones for the LexA-BD plasmid --> useable for Miniprep
    • counting: 
 - dh5a (Mariel) + pxj - KCM : 10^5 cfu/µg
 - dh5a (Mariel) + pxj - HS  : 4*10^4 cfu/µg
 - dh5a          + pxj - HS  : 1.5*10^5 cfu/µg
 - Top10         + pxj - HS  : 3.2*10^5 cfu/µg
 - db3.1         + pxj - HS  : 3*10^4 cfu/µg
  • The new BioBricks arrived so we made an overnight culture of, in LB(Amp):
    • p1010 in pSB1A3
    • BBa_c0012 (lacI)
    • BBa_c0051 (lamda cI)
    • BBa_j63001 (EYFP)
  • We plated p1010 in pSB3K3 on LB Kan
  • Digestion of the plasmid of the BBa_J63002(?) clone with XbaI and SpeI
    • 1x double digestion
    • 1x step by step digestion
    • result:
      • digestion seem to have work out in both cases
      • but the gel shows a high contamination with low molecular RNA, so we know now that RNase is crutial in the eluation buffer
      • the identity of the clone is still not verifified, because the size small fragement could be determinated
      • but as the new BioBricks arrived today, there is no longer an interest to go on working on this clone

General

  • Modeling meeting with Mr. Haich

Quote of the day

Marius loves Michis Kidneys!

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