Template:Team:KULeuven/28 August 2009/BlueLightReceptor

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# miniprep of liquid cultures of pSB3K3 grown overnight.  
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# Miniprep of liquid cultures of {{kulpart|pSB3K3}} grown overnight.  
{| border ="1" align="center"
{| border ="1" align="center"
!| Part || concentration (ng/μl) || 260/280 λ ||
!| Part || concentration (ng/μl) || 260/280 λ ||
|- align="center"
|- align="center"
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| pSb3K3 1 || 36,4 || 2,23 ||  
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| {{kulpart|pSB3K3}} 1 || 36,4 || 2,23 ||  
|- align="center"
|- align="center"
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| pSb3K3 2 || 33,0 || 2,46 ||
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| {{kulpart|pSB3K3}} 2 || 33,0 || 2,46 ||
|}
|}
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# gel electroforesis of pSB3K3 miniprep and of restriction digest from 27/08 ({{kulpart|BBa_J23101}} and {{kulpart|BBa_E0240}}. There was an unexpected signal at 800bp at the ({{kulpart|BBa_J23101}} lanes. the conclusion was that this part can not be used and we have to start all over from the basic DNA to make this ligation C.
+
# Gel electrophoresis of {{kulpart|pSB3K3}} miniprep and of restriction digest from 27/08 ({{kulpart|BBa_J23101}} and {{kulpart|BBa_E0240}}. There was an unexpected signal at 800bp at the ({{kulpart|BBa_J23101}} lanes. The conclusion was that this part can not be used and we have to start all over from the basic DNA to make this ligation C.
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# we checked our cultures that were radiated with blue light on 27/08.  
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# We checked our cultures that were radiated with blue light on 27/08.  
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#*the liquid cultures were put in the FACs machine. there was some fluorecense which is probably due to GFP. however, it can not be excluded that this is due to leakage from our promotor. so a new set up needs to be made where decent controls are included.  
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#*The liquid cultures were put in the FACs machine. There was some fluorescence which is probably due to GFP. However, it can not be excluded that this is due to leakage from our promotor. So, a new set up needs to be made where decent controls are included.  
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#*the LB plates were put under a special GFP lamp. Just like the liquid cultures there was signal but the same considerations need to be made.  
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#*The LB plates were put under a special GFP lamp. Just like the liquid cultures, there was signal but the same considerations need to be made.  
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# a new plan was deviced in order to make a permanent glycerol stock of LigA and to make a new set up for GFP measurements, this time including controls.  
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# A new plan was designed in order to make a permanent glycerol stock of LigA and to make a new set up for GFP measurements, this time including controls.  
#* LigA and {{kulpart|BBa_E0240}} were electroporated, plated and put overnight in the 37°C incubator.
#* LigA and {{kulpart|BBa_E0240}} were electroporated, plated and put overnight in the 37°C incubator.
#* {{kulpart|BBa_E0240}} was also plated starting from a glycerol stock and grown overnight at 37°.
#* {{kulpart|BBa_E0240}} was also plated starting from a glycerol stock and grown overnight at 37°.
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the following was done over the weekend:
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The following was done over the weekend:
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*saterday:
+
*Saturday:
** LigA grew well overnight. 4 colonies were selected for further testing and plated on new plates
** LigA grew well overnight. 4 colonies were selected for further testing and plated on new plates
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** the plates with {{kulpart|BBa_E0240}} overgrew so a single colony was picked and replated  
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** The plates with {{kulpart|BBa_E0240}} overgrew, so a single colony was picked and replated  
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**the plates containing electroporated ligC from 26/08 had two colonies after all. these were ented in liquid cultures and grown overnight so it can be checked later if these colonies do in fact contain ligC.
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** The plates containing electroporated ligC from 26/08 had two colonies after all. These were ented in liquid cultures and grown overnight, so it can be checked later if these colonies do in fact contain ligC.
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*sunday:
+
*Sunday:
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**the 4 colonies that were selected on saterday grew well and were ented in liquid cultures.
+
** The 4 colonies that were selected on Saturday grew well and were ented in liquid cultures.
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** some colonies from {{kulpart|BBa_E0240}} were also ented in liquid culture.
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** Some colonies from {{kulpart|BBa_E0240}} were also ented in liquid culture.

Latest revision as of 09:30, 8 September 2009

  1. Miniprep of liquid cultures of grown overnight.
Part concentration (ng/μl) 260/280 λ
1 36,4 2,23
2 33,0 2,46
  1. Gel electrophoresis of miniprep and of restriction digest from 27/08 ( and . There was an unexpected signal at 800bp at the ( lanes. The conclusion was that this part can not be used and we have to start all over from the basic DNA to make this ligation C.
  2. We checked our cultures that were radiated with blue light on 27/08.
    • The liquid cultures were put in the FACs machine. There was some fluorescence which is probably due to GFP. However, it can not be excluded that this is due to leakage from our promotor. So, a new set up needs to be made where decent controls are included.
    • The LB plates were put under a special GFP lamp. Just like the liquid cultures, there was signal but the same considerations need to be made.
  3. A new plan was designed in order to make a permanent glycerol stock of LigA and to make a new set up for GFP measurements, this time including controls.
    • LigA and were electroporated, plated and put overnight in the 37°C incubator.
    • was also plated starting from a glycerol stock and grown overnight at 37°.

The following was done over the weekend:

  • Saturday:
    • LigA grew well overnight. 4 colonies were selected for further testing and plated on new plates
    • The plates with overgrew, so a single colony was picked and replated
    • The plates containing electroporated ligC from 26/08 had two colonies after all. These were ented in liquid cultures and grown overnight, so it can be checked later if these colonies do in fact contain ligC.
  • Sunday:
    • The 4 colonies that were selected on Saturday grew well and were ented in liquid cultures.
    • Some colonies from were also ented in liquid culture.
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