light cycler cycle times - (Apr/16/2008 )
what kind of light cycler cycle times does everyone use??? i usually use an iCycler but have moved to a new lab where the lightcycler kind of rules... i was shocked to see that they use the following cycle times
denature - 1 second
anneal - 10 seconds
extend - 15 seconds
obviously this is very quick (and it seems to work) but when i spoke to the supplier of my mastermix they said this was strange???
I presume you're using the lightcycler with glass capillaries. Those times are fine. In fact, you could get away with 0 second pauses at the denature and anneal temps.
If your cycler uses plastic tubes, you'll possibly need to extend the times. This month's issue of Biotechniques (www.biotechniques.com) has a couple of papers comparing plastic and glass in fast cyclers, as well as the efficiency of thermocyclers. Registration is free, and the journal does have plenty of cool protocols.
I am not sponsored by Biotechniques, nor are the papers mine.
Yep, as swanny said, these times sound more than fine for Lightcycler (actually, they might be a bit too long for certain transcripts!). That's why I used to love the LC so much!!! You would get your results, melting curve included, in ~45min!