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Running gel after RT-PCR


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5 replies to this topic

#1 Ahrenhase

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Posted 07 April 2009 - 10:32 AM

Say I wanted to run a gel after finishing an RT-PCR SYBR green reaction, would I have to restain or would the SYBR in this master mix be enough the see?

#2 dvddecarvalho

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Posted 07 April 2009 - 11:20 AM

I usually run agarose gel after my qRT-PCR SYBR reactions.
I stain it with etbr. I`m not sure if we can see SYBR stain in UV aparatus.

#3 mad

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Posted 07 April 2009 - 10:28 PM

I'm not sure if it helps but usually in my lab everyone just stains the gel with etbr

better to be safe than sorry i guess unless you are very certain the sybr will light up

MAD :wacko:

#4 mtrnbh

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Posted 07 April 2009 - 10:48 PM

we also use etbr for staining the SYBR RT-PCR products.

#5 almost a doctor

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Posted 08 April 2009 - 04:24 AM

Say I wanted to run a gel after finishing an RT-PCR SYBR green reaction, would I have to restain or would the SYBR in this master mix be enough the see?


that will actually depend on the sensitivity of your imaging equipement, so you might want to add more SYBR, or just EtBr as suggested, to make sure.

Alternatively, you can run your samples, check for SYBR, and if you cant see them, then stain the gel with EtBr post-electrophoresis.

#6 Ahrenhase

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Posted 08 April 2009 - 11:10 AM

I think it would fluoresce, but if I'm bored enough tomorrow I'll try it out and let you know how it works out.

yeah, if it doesn't show up I can just stain it afterwards anyway in more SYBR green.




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