IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/29

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Results of transformation with our ligation products

  • Everything grew! Better efficiency with electrop. 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 (with cutting sites) expected size after PCR (about)
Pupp 255 480
Pspac 125 350
RBS S 56 280
RBS W 56 280
agrD 200 430
  • Result : nothing, even no ladder, problem with the gel!

- run again on a e-gel

  • Lane1 :ladder 100bp
  • Lane2 : Pupp colony 1
  • Lane3 : Pupp colony 3
  • Lane4 : Pspac colony 1
  • Lane5 : Pspac colony 3
  • Lane6 : RBS S colony 1
  • Lane7 : RBS S colony 4
  • Lane8 : RBS W colony 1
  • Lane9 : RBS W colony 4
  • Lane10 : agrD colony 1
  • Lane11 : agrD colony 4
  • Lane12 : HyperladderI
  • Result : nothing, just the primers! Problem with our PCR

Plate biobricks from MIT

  • E0840
  • B0014
  • I712007
  • C0012
  • B0015
  • C0061
  • R0063


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