Problems with RNA extraction or QPCR for AI

[deki]

Can somebody help me,
When I test samples (alantoic fluid) on Avian Influenza, using QIAamp Viral RNA Kit for extraction and Taqman Influenza A/H5 detection kit version 1.0 for QPCR detection, I have no problem.
I am having problems with RNA extraction or Q-RT-PCR when I test samples that are inactivated with formaldehyde. The results are always negative, and when i test the same sample without inactivation fith formaldehyde, results are positive. Where is the problem, in extraction or in Q-RT-PCR reaction?

[MARGARITA MARTINEZ]

Hello Deki.

The Problem You Have Is Same For Me.

I Think The Inactivation Whit Formaldehydo
To Spoil The Core Of Genoma Viral And Estruture Of Rna....

[danfive]

Formaldehyde is a fixative, you've crosslinked your RNA (RNA-RNA, RNA-Protein). Carrying over formaldehyde to your PCR reaction will also crosslink your enzymes (protein). I've heard of reverse crosslinking with high salt incubation see [Only registered users see links. ]

[danfive]

Originally Posted by danfive

Formaldehyde is a fixative, you've crosslinked your RNA. Carrying over formaldehyde to your PCR reaction will also crosslink your enzymes (protein). I've heard of reverse crosslinking with high salt incubation see [Only registered users see links. ]

See [Only registered users see links. ] for effects of formaldehyde on RNA.