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