Daniel:Notebook/ComboLock/2017-3-6
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Padlock0601 Test (Started Wed March 1)
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
- Maximal Rolling Circle Amplification
- Add 15 uL master mix to each tube
- Add 4 uL template according to table
- Incubate at 95 C for 5 minutes
- Lower to 55C for 15 minutes
- Lower to 30C for at least 2 minutes
- Add 1 uL Phi29 polymerase to each sample
- Incubate 15 hours at 30C
- Heat kill enzyme by incubating 10 minutes at 65C; Hold at 12C