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Post by xiaolong88 » Tue Dec 11, 2012 4:58 pm


I have transcript (RNA) which I would use for qRT PCR for standard curve-the standard.
I should quantitate the transcript (RNA) before reverse transcribe them into cDNA.
I wonder I should serially dilute the transcript and perform reverse transcription? Or I should synthesis the cDNA first and only perform serial dilution for qPCR? Any different? :roll:

Hope you could help me :)

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Post by JackBean » Tue Dec 11, 2012 11:00 pm

you should first make the cDNA as soon after RNA extraction as possible and then you can handle it as you want ;)

Cis or trans? That's what matters.

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Post by bravebeaker » Wed Dec 26, 2012 4:47 am

1. extracted RNA --> DNase treatment --> RT to synthesize cDNAs
2. use same primers to amplify your target by PCR
3. extract bands from gel and purify target DNA
4. use the purified DNA for making a standard curve.
5. runa a qPCR
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