Can I detect any amplicon less than 50bps with qPCR?

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Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

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Seyed Elyas Meshkani

Hello,

Yes, it's applicable. Also, you can use gyrB as an internal control.

Make sure to eliminate probable primer-dimer.

Furthermore, you can shorten the extension time.

Best,

Ed Hamdeh

You can but if you wanted a more optimal approach I'd suggest using a modified oligo(dT) technique. Way quicker and only need 1 RT reaction. I have the paper somewhere I'll have to look. Message or email me a reminder tomorrow if youre interested. [email protected]

Yasu Xu

Hi,

I do not think I have a polyA tail for those crRNA I want to sequence.

Hi Elyas,

Thanks! Also, I noticed people recommend SYBR GREEN for 100-200bps and Taqman for shorter amplicon. Have you ever tried some shorter sequence qPCR experiment?

Dear Yasu,

I would recommend SYBR because TaqMan results in later CTs.

Seyed Elyas Meshkani Okay! Thanks!