User:Daniela A. Garcia S./Notebook/Modeling UNAM-Genomics Mexico/2010/08/03

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Wet Lab: OmpC/F promoter

OmpC transformation

The part [BBa_R0082] (pOmpC) inserted in pSB1A2 was transformed in DH5α competent cells with the following protocol. The plasmid N.19 ([BBa_I51020]) was used as positive control.


  • 5 μL of BBa_R0082
  • 5 μL plasmid N. 19


1) 20' on ice

2) 1' 42ºC

3) 5' on ice

4) 60' in 1 ml of LB medium at 37ºC

5) 1' at 13000 rpm


Two petri boxes were made with LBam to plaque the transformed cells. 100 μL were added and left overnight at 37ºC.

Electrophoresis Gel OmpF PCR_RTTH

In order to probe the PCRs reactions an electroforesis gel was made. This was loadded as follows:

  • Lane1. Ladder
  • Lane2. B3K3 OmpF
  • Lane3. B3K3 control
  • Lane4. B1T3 OmpF
  • Lane5. B1T3 control








Note

The relation for all the antibiotics used is 1:1000

Modeling: BioBrick promoters

From the article: "Measuring the activity of BioBrick promoters using an in vivo reference standard"

  • Rate of transcription initiation as the property to be measured. Promoter clearance rate as the specific property that best describes transcription initiation (Promoter Activity )



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