User:Daniela A. Garcia S./Notebook/Modeling UNAM-Genomics Mexico/2010/08/02

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Wet Lab: OmpC/F promoter

OmpC obtaining

The OmpC promoter was obtained from the 2010 Plate [Plate 1_ Well K16]. The part is BBa_R0082 and is inserted in pSB1A2 [A=Ampicillin]

  • 15μl HPLC were added

OmpF PCR

PCR1.

Primer forward: Suffix Primere reverse: Bluepromoter-Preffix Template: pSB3K3

PCR2.

Primer forward: Suffix Primere reverse: OmpFpart1-Preffix Template: pSB3K3

PCR3.

Primer forward: Suffix Primere reverse: Bluepromoter-Preffix Template: pSB1T3

PCR4.

Primer forward: Suffix Primere reverse: OmpFpart1-Preffix Template: pSB1T3


  • General PCR mixture:
    • 4 microL Template(dilution 1:4)
    • 3 microL Primer forward (5mM)
    • 3 microL Primere reverse (5mM)
    • 6 microL HPLC
    • 6 microL 3.3X XL Buffer
    • 3 microL Mg solution
    • 5 microL dNTP's (0.4 mM)


  • Enzyme(RTTH) mixture for PCR Reaction:
    • 10.5 microL HPLC
    • 9 microL 3.3X XL Buffer
    • 0.5 microL RTTH


  • Program [35 PCR cycles]
    • 94ºC 5min
      • 94ºC 45seg
      • 60ºC-55ºC 45seg
      • 72ºC 10min
    • 72ºC 10min
    • 4ºC 3min