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Copy number assay using Real time Pcr-SYBR green assay?


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#1 GNANA

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Posted 21 September 2010 - 12:49 PM

Hi everybody,
I am again here expecting ur guidance for my current task of doing copy number assay using DNA in Real time PCR-SYBR green. i got to screen the copy number of my gene of interest in various cell lines, could anyone please suggest and explain me how to analyse the result and what method. am going to use a control Dna from normal cell line and b-globulin as an internal control. i was asked to do it by delta ct method, but i dnt know whether it s the correct method to do for copy number also. if dct method is the right one can i analyse the data with the same way i do for expression?..thanks in advance for any of ur suggestions....

gnana..
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#2 Darktan

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Posted 22 September 2010 - 03:23 AM

Why not using commercially available TaqMan copy number assays?
(could be much easier)
If that is not an option, I encourage you to read about copy number assays by the AB. There you could find the answers to your questions.

#3 Trof

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Posted 22 September 2010 - 03:24 AM

You have reference (b-globulin) and you have calibrator (normal cell line), you just use delta-delta Ct as usual.

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#4 GNANA

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Posted 22 September 2010 - 03:51 AM

thanks trof & Darktan..i cannot ask my prof to order for taqman now,so had to go with SYBR only..i do the assay and get back to you if i had any queries....

gnana...

Edited by GNANA, 22 September 2010 - 03:54 AM.

I would prefer being perfectionist rather than a passionist in Research.

I always had an alternate hypothesis....




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