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Template:Team:KULeuven/27 August 2009/BlueLightReceptor
From 2009.igem.org
(Difference between revisions)
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#** {{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 | ||
| - | # 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 | + | # 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 | ||
Revision as of 14:23, 27 August 2009
- the electroporations of 26/08 were checked.
- pSB3K3 had some colonies. they are ented in liquid culture today.
- LigC ( + ) did not grow. we figured that an insert of 35bp was to short to ligate so we decided to use as insert in stead of 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
