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SYBR vs FAM probe

RT-PCR

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#1 HOYAJM

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Good

Posted 13 November 2012 - 12:24 PM

I will be looking at virus replication by real-time PCR using primers for antigenomic RNA. I originally wanted to do this with a FAM probe for increased sensitivity, but I am having much better luck with SYBR green.

I am unfamiliar with what is acceptable for measuring virus replication. If I was to present my data measuring replication by RT-PCR with SYBR green, would reviewers question why I did not use a more sensitive technique (i.e. FAM probe)?

I am hoping both are OK, but I was just wondering if anybody has published similar data using SYBR green with no problems.

Thanks,

Jared

#2 Trof

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Excellent

Posted 13 November 2012 - 03:33 PM

Both types of methods are AFAIK accepted equally for quatitative real-time PCR, the probe increases the specificity, but with SYBR you can (and you should) do always a melting analysis after run to chcek wheter you have the right product.
I think SYBR with melting curve will be OK.

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