IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/29: Difference between revisions

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- Load a gel (1.3% agarose) : 5μL of PCR products + 1μL of dye (only 1μL of 100b ladder)
- Load a gel (1.3% agarose) : 5μL of PCR products + 1μL of dye (only 1μL of 100b ladder)
In the death plasmid, the VF-VR size is about 280b.


{|class="wikitable" style="text-align:center" border="1"
{|class="wikitable" style="text-align:center" border="1"
Line 41: Line 43:
!  Transformed products !! size of the product !! expected size after PCR  
!  Transformed products !! size of the product !! expected size after PCR  
|-
|-
| Pupp || 2 || 2 || 0
| Pupp || 255 || 535
|-
|-
| Pspac || 2 || 2 || 0
| Pspac || 125 || 405
|-
|-
| RBS S || 3 || 0 || 2
| RBS S || 56 || 336
|-
|-
| RBS W || 3 || 0 || 2
| RBS W || 56 || 336
|-
|-
| agrD || 3 || 2 || 0
| agrD || 200 ||280
|}
|}
* Result : nothing, even no ladder, problem with the gel!
- run again on a e-gel
*Result : nothing, just the primers! Problem with our PCR


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Revision as of 03:39, 1 September 2008

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Results of transformation with our ligation products

  • Everything grew! Better efficiency with electro> than with chemical protocol

Single colony PCR to check our transformation

Transformed products number of picked colonies from chemical transformation number of picked colonies from electrop. transformation (neat) number of picked colonies from electrop. transformation (1:10)
Pupp 2 2 0
Pspac 2 2 0
RBS S 3 0 2
RBS W 3 0 2
agrD 3 2 0

- Single colony PCR : 13μL of SDW+cells, 5μL of Master Mix, 1μL of VF and 1μL of VR (and plate each single colony)

- Load a gel (1.3% agarose) : 5μL of PCR products + 1μL of dye (only 1μL of 100b ladder)

In the death plasmid, the VF-VR size is about 280b.

Transformed products size of the product expected size after PCR
Pupp 255 535
Pspac 125 405
RBS S 56 336
RBS W 56 336
agrD 200 280
  • Result : nothing, even no ladder, problem with the gel!

- run again on a e-gel

  • Result : nothing, just the primers! Problem with our PCR


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