IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/09/02: Difference between revisions

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* Glycerol stocks (about 20 tubes of each)
* Glycerol stocks (about 20 tubes of each)


==Check transformation in E.coli of our ligated products==


- Single colony PCR with VF and VR : add 10μL of SDW, 5μL of MM, 1μL of VF, 1μL of VR and 3μL of cells
{|class="wikitable" style="text-align:center" border="1"
|-
! inserted products !! number of colonies to check (chemical transf.) !! number of colonies to check (electrop. transf.)
|-
| RBS S || 3 || 2
|-
| RBS W || 3 || 2
|-
| Pspac || 2 || 2
|-
| Pupp || 2 || 2
|-
| agrD || 3 || 2
|-
|}
- Gel (3% of agarose)


==Check transformation in E.coli of our ligated products==
==Check transformation in E.coli of our ligated products==

Revision as of 02:10, 4 September 2008

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Transformation of Bacillus

  • with fresh plates from yesterday, last protocol
Time (min) 0 20 40 60 85 105 125
OD650 for IA751 0.1192 0.1330 0.1736 0.2238 0.3124 0.4523 0.5113
OD650 for IA771 0.1144 0.1216 0.1593 0.2296 0.3476 0.4302 0.5565
  • t0 (IA751) = 100min
  • t0 (IA771) = 110min
  • Transformation

- IA751 on Cm5 and Spc 50 (control DNA less plates)

- IA751 + ECE112 (1μg of DNA) on Cm5

- IA751 + ECE153 (0.6μg of DNA) on Scp50

- IA771 on Cm5 (control)

- IA771 + ECE166 on Cm5

  • Glycerol stocks (about 20 tubes of each)


Check transformation in E.coli of our ligated products

- Single colony PCR with VF and VR : add 10μL of SDW, 5μL of MM, 1μL of VF, 1μL of VR and 3μL of cells

inserted products number of colonies to check (chemical transf.) number of colonies to check (electrop. transf.)
RBS S 3 2
RBS W 3 2
Pspac 2 2
Pupp 2 2
agrD 3 2

- Gel (3% of agarose)


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