Template:Team:KULeuven/27 August 2009/BlueLightReceptor

From 2009.igem.org

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# the electroporations of 26/08 were checked.  
+
# The electroporations of 26/08 were checked.  
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#*pSB3K3 had some colonies. they were ented in liquid culture.
+
#*{{kulpart|pSB3K3}} had some colonies. they were ented in liquid culture.
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#*LigC ({{kulpart|BBa_J23101}} + {{kulpart|BBa_E0240}}) did not grow. we figured that an insert of 35bp was to short to ligate so we decided to use {{kulpart|BBa_E0240}} as insert and {{kulpart|BBa_J23101}} + pSB1A2 as vector . The following restriction digest was started:
+
#*LigC ({{kulpart|BBa_J23101}} + {{kulpart|BBa_E0240}}) did not grow. we figured that an insert of 35bp was too short to ligate so we decided to use {{kulpart|BBa_E0240}} as insert and {{kulpart|BBa_J23101}} + {{kulpart|pSB1A2}} as vector . The following restriction digest was started:
#** {{kulpart|BBa_J23101}} was cut with SpeI and PstI
#** {{kulpart|BBa_J23101}} was cut with SpeI and PstI
#** {{kulpart|BBa_E0240}} was cut with XbaI and PstI   
#** {{kulpart|BBa_E0240}} was cut with XbaI and PstI   
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# the miniprep that was made to sequence the LigA construct on 26/08 was used again to perform a restriction digest (EcoRI and PstI). this was put on gel to check wether there actually was LigA-insert in the vector and wether the insert had the right length. the gel showed a signal at 1000 bp and at 2000 bp which coincides with the insert (BLp + GFP) and the vector.
+
# The miniprep that was made to sequence the LigA construct on 26/08 was used again to perform a restriction digest (EcoRI and PstI). This was put on gel to check whether there actually was LigA-insert in the vector and whether the insert had the right length. The gel showed a signal at 1000 bp and at 2000 bp which coincides with the insert (BLp + GFP) and the vector.
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# a new set up to light the Ecoli was engineered.  
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# A new setup to light the E.coli was engineered.  
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#* fresh cultures were made from the old ones (LB plate ligA 14/08 and the two liquid cultures from 26/08 were used as templates.)
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#* Fresh cultures were made from the old ones (LB plate ligA 14/08 and the two liquid cultures from 26/08 were used as templates.)
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#* they were put in the 16°C room for about an hour
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#* They were put in the 16°C room for about an hour
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#* a blue light (40W) was put on them for about an hour
+
#* A blue light (40W) was put on them for about an hour
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#* they were put in the 37°C incubator overnight
+
#* They were put in the 37°C incubator overnight

Latest revision as of 09:24, 8 September 2009

  1. The electroporations of 26/08 were checked.
    • had some colonies. they were ented in liquid culture.
    • LigC ( + ) did not grow. we figured that an insert of 35bp was too short to ligate so we decided to use as insert and + as vector . The following restriction digest was started:
      • was cut with SpeI and PstI
      • was cut with XbaI and PstI
  2. The miniprep that was made to sequence the LigA construct on 26/08 was used again to perform a restriction digest (EcoRI and PstI). This was put on gel to check whether there actually was LigA-insert in the vector and whether the insert had the right length. The gel showed a signal at 1000 bp and at 2000 bp which coincides with the insert (BLp + GFP) and the vector.
  3. A new setup to light the E.coli was engineered.
    • Fresh cultures were made from the old ones (LB plate ligA 14/08 and the two liquid cultures from 26/08 were used as templates.)
    • They were put in the 16°C room for about an hour
    • A blue light (40W) was put on them for about an hour
    • They were put in the 37°C incubator overnight
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