My experiment was to set up an efficiency curve for a gene using MCF-7 breast cancer cell line sample cDNAs. However, my amplification curves do have the shape of a correct amplification curve (Ct values also seem correct) but are jagged or zig-zag throughout and even recognized as 'negative' despite neat and specific corresponding melting curves. This is actually a second repeat of the experiment and the first repeat yielded ideal results. Therefore, I am wondering if there is a possible mechanical problem with the machine (eg. alignment of the xenon lamp). The machine I am using is Roche LightCycler 480.
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Trof
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Posted 13 April 2012 - 01:46 PM
You did use your plate multiple times, didn't you?
That happens. No more than two runs of one plate, and even the second can be a little bit zig-zagged.
Try the High specifity setting in the 2nd Derivative analysis module, it should make correct Cts.
That happens. No more than two runs of one plate, and even the second can be a little bit zig-zagged.
Try the High specifity setting in the 2nd Derivative analysis module, it should make correct Cts.
Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.
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