IGEM:UNAM-Genomics-Mexico/2009/Notebook/iGEM 2011 UNAM-Genomics Mexico/2011/06/10

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Transformation of the PBBRMCS-5 and the PCR amplification of PSB1T3 ligation into DH5α

The ligation of PBBRMCS-5 and the PCR amplification PSB1T3 transformed into E. coli DH5α

Transformation

1 tube with 100 μL of competent E. coli DH5α cells with 8 μL of the PBBRMCS-5 and PSB1T3 ligation 1 tube with 50 μL of competent E. coli DH5α cells with no DNA as a control.

  1. Leave the tubes for 20 minutes in ice.
  2. Leave 1 minute at 42ºC for a Heat Shock.
  3. Leave for 5 minutes in ice.
  4. Add 1 mL of liquid LB.
  5. Incubate at 37ºC during 1 hour.
  6. Plate in Petri Dishes with 30 mL of lb solid and gentamicin each petri dish and 200 μL of the transformation in every dish.

Incubate over night and check for red colonies the next day.