Daniel:Notebook/ComboLock/2017-3-6

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Revision as of 00:13, 7 March 2017 by >Djacobse (Created page with "=Padlock0601 Test (Started Wed March 1)= Back to Calendar After reading some papers I'm going to give it...")
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Padlock0601 Test (Started Wed March 1)

Back to Calendar

After reading some papers I'm going to give it one more go to see if I can't amplify the DNA from last week better. I'll use the method of largest amplification I know: most time using random hexamers as primers.

Protocol

  1. Maximal Rolling Circle Amplification
    1. Add 15 uL master mix to each tube
    2. Add 4 uL template according to table
    3. Incubate at 95 C for 5 minutes
    4. Lower to 55C for 15 minutes
    5. Lower to 30C for at least 2 minutes
    6. Add 1 uL Phi29 polymerase to each sample
    7. Incubate 15 hours at 30C
    8. Heat kill enzyme by incubating 10 minutes at 65C; Hold at 12C