Template:Team:KULeuven/27 August 2009/BlueLightReceptor

(Difference between revisions)

Revision as of 14:25, 27 August 2009

the electroporations of 26/08 were checked.
- pSB3K3 had some colonies. they were ented in liquid culture.
- LigC ( + ) did not grow. we figured that an insert of 35bp was to short to ligate so we decided to use as insert and + pSB1A2 as vector . The following restriction digest was started:
  - was cut with SpeI and PstI
  - was cut with XbaI and PstI
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.
a new set up to light the Ecoli 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

@@ Line 1: / Line 1: @@
 # the electroporations of 26/08 were checked.
-#*pSB3K3 had some colonies. they are ented in liquid culture today.
+#*pSB3K3 had some colonies. they were ented in liquid culture.
-#*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 in stead of 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 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:
 #** {{kulpart|BBa_J23101}} was cut with SpeI and PstI
 #** {{kulpart|BBa_E0240}} was cut with XbaI and PstI