User:Jorge Arturo Zepeda/Notebook/iGem LCG-UNAM team 2010/2010/04/19

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Monday 19th April 2010

Heat shock transformation:

  • Take 50ul of competent cells from Miguel’s lab frozen at -70ºC, out of a 100ul eppendorf into a new one (sterilized), then I add 5ul of the DNA (luciferase from Photinus pyralis BBa_I712019) and put into ice for 20 minutes.
  • After that give it a heat shock at 42 ºC for one minute, then put it in ice for another 5 minutes.
  • Now grow it in 1ml of liquid LB medium with no antibiotic and shaking for one hour.
  • Then centrifuge for 1 minute at 13 rpm and keep the pellet with 100ul, now plate those 100ul in a petri box with kanamycin using the triangle, also plate a control with the sole medium and put them in the incubator at 37ºC overnight.