Trof, on 13 May 2011 - 02:16 AM, said:
As I understand it, you don't really isolate DNA, you just lyse the cells and then do PCR?.
Yes, that's the way it's done. About 5 colonies are taken off the plates and brought into dH2O. There is no purification prior to the PCR. I was too quick in my explanation above.
gebirgsziege, on 13 May 2011 - 04:04 AM, said:
Suspending the cells will not lyse them, the lysis of the cells happens in two steps during the protocol you described: (i) by freezing your samples you create ice crystals in the bacterial cells which will destroy some of the cells (i.e. ice has a larger volume than water). This facilitates the lysis of the cells prior to (ii) cracking them open by "cooking" them in the PCR.
If you plan to use your samples more than one/two times you should think about something to stabilise the DNA as suggested in the previous post.
Okay, thank you for your answer! I will implement that in my study report.
Thanks for all the quick replies