Copy number

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danush's picture
Copy number

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.


Amtekoth's picture
If you are looking for at

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?