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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