Biology Forum › Molecular Biology › qPCR
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- December 11, 2012 at 4:58 pm #17133
xiaolong88ParticipantHi,
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? 🙄Hope you could help me 🙂
- December 11, 2012 at 11:00 pm #113227JackBeanParticipant
you should first make the cDNA as soon after RNA extraction as possible and then you can handle it as you want 😉
- December 26, 2012 at 4:47 am #113300bravebeakerParticipant
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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