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E. coli transformation: Difference between revisions
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* Took 1 colony from PUC19 dh5a plate and mixed with 250ul RO water | * Took 1 colony from PUC19 dh5a plate and mixed with 250ul RO water | ||
* Incubated at 98C for 15 minutes, then hold at 10C (boiling program in PCR machine) - to lyse the cells | * Incubated at 98C for 15 minutes, then hold at 10C (boiling program in PCR machine) - to lyse the cells | ||
* Put on 2 PCRs on Col program (53C annealing) - one with M13 primers (for PUC19), one with S16 primers (for any bacteria), then hold for 4C for 20 hours. | * Put on 2 PCRs on Col program (53C annealing) - one with M13 primers (for PUC19), one with S16 primers (for any bacteria), then hold for 4C for 20 hours. (Sam said 56 might be better for M13 but is too hot for s16) | ||
* From top - ladder, M13, S16 | * From top - ladder, M13, S16 | ||
