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danush

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Hi, I am trying to quantify copy number of a gene. I have aquestion. Which is the best method, standar curve with a calibrator, Delta-Delta Ct? The calibrator for the standard curve could be the gene cloned in a plasmid (only one copy)? Do I have to use a housekeeping (I assume yes).
Thanks for the help.

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Posted Nov 15, 2007, 19:16 PM
Amtekoth

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If you are looking for at cellular DNA, as opposed to RNA, then I should think that any gene known to be unamplified (2 copies per diploid cell) should work, as long as your primers are specific for that sequence. Then either a plasmid or an amplicon of that sequence should work as a calibrator.

I think?

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"God put me on this earth to accomplish a certain number of things. Right now I am so far behind that I will never die."- Bill Watterson

Posted Dec 11, 2007, 7:16 AM
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