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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. | ||
| - | #*pSB3K3 had some colonies. they were ented in liquid culture. | + | #*{{kulpart|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 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 to 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 | ||
Revision as of 11:57, 1 September 2009
- 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 to 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
- 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
