definition IPC and offset

Here we discuss the use and processing of inter-plate calibrators

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definition IPC and offset

Postby Jeroen » Mon Nov 07, 2011 6:02 pm

Dear Sir/Madam,

During the course "qPCR data analysis" you showed some rather nice schemes describing the necessity for IPC in a multiplate study. You also showed that there is no need to incorporate the offset (in a multiplate study, if all genes are analyzed for each sample in the same plate or if all samples are analyzed for each gene in the plate). Do I understand it correctly that there is no need for including the offset since this is incorporated in the IPC? And could you please give me a definition for the offset, just to make sure I have a correct understanding of the definition?

My apologies for this rather basic question.

Regards,
Jeroen
Jeroen
 
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Re: definition IPC and offset

Postby Mikael Kubista » Mon Nov 07, 2011 9:40 pm

Offset is a systematic difference between the measured values in a plate and an arbitrary base level. The offset is in general not known.

You can easily verify if you need IPC or not. Merge data from two plates and pre-process them without using IPC. Then start over merge the data again, but this time add one cycle to all Cq in one plate. If this affects the result of pre-processing you need IPC; if it has no effect, IPC is not needed.

Good luck
Mikael Kubista
 
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Joined: Tue Jul 01, 2008 12:28 pm


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