IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/09/10

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PCR of GFP+RBS and Promoter+RBS
- PCR

- Run a 1.2% agarose gel with 1μL of sample


 * Lane 3 : HyperladderI
 * Lane 4 : GFP + RBS 1A
 * Lane 5 : GFP + RBS1B
 * Lane 6 : GFP + RBS 2A
 * Lane 7 : GFP + RBS 2B
 * Lane 8 : Pupp + RBS 1
 * Lane 9 : Pupp + RBS 2
 * Lane 10: HyperladderIV


 * Result



RBS screening

 * 5 colonies from chemical transformation + 6 colonies from electroporation transformation for RBS S
 * 5 colonies from chemical transformation + 6 colonies from electroporation transformation for RBS W

- PCR : 11μL of SDW + 5μL of MM + 1+1μL of primers (RBS detect + VR) + 2μL of cells (program iGEM34)

- Gel1


 * Lane2 : RBSS1
 * Lane3 : RBS S2
 * Lane4 : RBS S3
 * Lane5 : RBS S4
 * Lane6 : RBS S5
 * Lane7 : RBS S6
 * Lane8 : HyperladderIV
 * Lane9 : RBS S7
 * Lane10 : RBS S8
 * Lane11 : RBS S9
 * Lane12 : RBS S10
 * Lane13 : RBS S6 with VF and VR primers
 * Lane14 : RBS S11

- Gel2


 * Lane2 : RBSW1
 * Lane3 : RBS W2
 * Lane4 : RBS W3
 * Lane5 : RBS W4
 * Lane6 : RBS W5
 * Lane7 : RBS W6
 * Lane8 : HyperladderIV
 * Lane9 : RBS W7
 * Lane10 : RBS W8
 * Lane11 : RBS W9
 * Lane12 : RBS W10
 * Lane13 : RBS W6 with VF and VR primers
 * Lane14 : RBS W11


 * Result



- Nothing, except for the PCR with VF and VR primers, either our primers are not right, either we have no insert. But the gel is not good enough to see very well, we will try to run a new gel.


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