User:Daniela A. Garcia S./Notebook/Modeling UNAM-Genomics Mexico/2010/07/28

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Wet Lab: OmpF/C promoters characterization

Electrophoresis Gel OmpF PCR_RTTH

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

  • Ladder
  • positive control B3K3
  • B3K3 OmpF
  • positive control B1T3
  • B1T3 OmpF
OmpFPCR RTTH 270710.jpg

There were not positive results for none of the reactions, even for the B1T3 OmpF two bands were obtained.

OmpF PCR_TaqDNApol

A second set of PCR rections was made, this time with the TaqDNApol enzyme.


PCR's Programme

  • 94ºC, 5:00
  • 35x
    • 94ºC, 0:45
    • 55ºC, 0:45
    • 72ºC, 3:00
  • 72ºC, 10:00 <<
  • 4ºC, ...

The temperature for extension (<<) was change. The one used for the RTTH PCR from 27/07/10 was to high, this cloud be a possible explanation why the reaction was unsuccesful.