Template:Team:KULeuven/26 August 2009/BlueLightReceptor

From 2009.igem.org

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# the plates with ligation A (blp + GFP) where fetched from the blue light installation. there was no GFP signal. the following actions will be taken:
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# The plates with ligation A (''blp'' + GFP) were fetched from the blue light installation. There was no GFP signal. The following actions will be taken:
-
#* the plasmids will be purified from the colonies and will be sequenced using primer 2260.
+
#* The plasmids will be purified from the colonies and will be sequenced using primer 2260.
-
#* next time we will probably put them in liquid cultures under the LEDs while shaking gently. also, other parameters that need to be considered are being researched.  
+
#* Next time, we will probably put them in liquid cultures under the LED's while shaking gently. Also, other parameters that need to be considered are being researched.  
-
#* they will not be exposed to the light as long anymore. we decided that 1h will be more than enough.
+
#* They will not be exposed to the light as long anymore. We decided that 1h will be more than enough.
-
#* possible bleaching?
+
#* Possible bleaching?
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# a colony from all three plates with the ligA-construct were taken and ented in liquid culture. this was needed to check wether the colonies were still alive.
+
# A colony from all three plates with the LigA-construct was taken and ented in liquid culture. This was needed to check whether the colonies were still alive.
#* culture 13/08
#* culture 13/08
#* culture 14/08 (1)
#* culture 14/08 (1)
#* culture 14/08 (2)  
#* culture 14/08 (2)  
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# by 5pm, one of the liquid cultures had grown and was miniprepped. 20μl was sent for sequencing together with 5μM of primer 2260.
+
# By 5pm, one of the liquid cultures had grown and was miniprepped. 20μl was sent for sequencing together with 5μM of primer 2260.
{| 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"
-
| ligA (14/08 (1)) || 23,1 || 2,21 ||
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| LigA (14/08 (1)) || 23,1 || 2,21 ||
|}
|}
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4. two electroporations were performed and plated on LB medium. one with ligC ({{kulpart|BBa_J23101}} + {{kulpart|BBa_E0240}}) and one with pSB3K3 DNA.
+
4. Two electroporations were performed and plated on LB medium. One with LigC ({{kulpart|BBa_J23101}} + {{kulpart|BBa_E0240}}) and one with {{kulpart|pSB3K3}} DNA.

Latest revision as of 12:49, 14 September 2009

  1. The plates with ligation A (blp + GFP) were fetched from the blue light installation. There was no GFP signal. The following actions will be taken:
    • The plasmids will be purified from the colonies and will be sequenced using primer 2260.
    • Next time, we will probably put them in liquid cultures under the LED's while shaking gently. Also, other parameters that need to be considered are being researched.
    • They will not be exposed to the light as long anymore. We decided that 1h will be more than enough.
    • Possible bleaching?
  2. A colony from all three plates with the LigA-construct was taken and ented in liquid culture. This was needed to check whether the colonies were still alive.
    • culture 13/08
    • culture 14/08 (1)
    • culture 14/08 (2)
  3. By 5pm, one of the liquid cultures had grown and was miniprepped. 20μl was sent for sequencing together with 5μM of primer 2260.
Part concentration (ng/μl) 260/280 λ
LigA (14/08 (1)) 23,1 2,21

4. Two electroporations were performed and plated on LB medium. One with LigC ( + ) and one with DNA.

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