IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/09/11

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(RBS screening)
Current revision (04:06, 16 September 2008) (view source)
(RBS screening)
 
Line 53: Line 53:
- Nothing, except for the PCR with VF and VR primers, either our primers are not right, either we have no insert.
- Nothing, except for the PCR with VF and VR primers, either our primers are not right, either we have no insert.
 +
==RBS screning but single digest==
 +
 +
- Grow some colonies of Top10 transformed with RBS into 10mL of LB with antibiotic
 +
 +
We want to check our transformation with single digest. If we have self ligation in our transformation, we will loose the XbaI cutting site.
 +
 +
==Backbone for ligation==
 +
 +
- New pellets in 600μL of SDW
 +
 +
- Plasmid miniprep
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RBS screening

- We run our PCR products from yesterday on a new 1.8% agarose gel

- 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.

RBS screning but single digest

- Grow some colonies of Top10 transformed with RBS into 10mL of LB with antibiotic

We want to check our transformation with single digest. If we have self ligation in our transformation, we will loose the XbaI cutting site.

Backbone for ligation

- New pellets in 600μL of SDW

- Plasmid miniprep


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