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problem with agarose gel? - (Jul/27/2005 )

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Hi everybody!
I was not on a vacation, but work really hard on my problem. I found the help - girl from the other lab. Nice girl smile.gif

So, We tried wlmost every combination what could be wrong. So we proved that my agarose is just fine and could not be the main problem. Also my electroforesis machine is fine. But now we are trying to prove, that is sth wrong with my SYBR Geen. 'cause when we used EtBr, my bands, with my agarose, with my EF machine, was just perfect !!!!!!! But when we used my SYBR Green and the same PCR products my "bands" were just like they on pictures. So today I am trying to prove once again that the problem is in dye.
Sorry I could not send you a picture, to compare the SBGreen and EtBr, 'cause we worked with this product in her lab(in our lab its forbidden to use EtBr) wich has not a digital camera.
I will repoort what is goping on, and hope that if someone else has the same problem, I could help him.

Thaks

-k4bf0389-

Here I am!
You couldn't believe. Here I sent to you two pictures. One picture with funny "bands" is dyed with SYBR Green(SG), the second with SYBR Safe(SS). Products are not the same, but everything else condition are the same. The difference is evident. I could not believe. So I decided, that sth is wrong with my SYBR Green, so I will not use it any more.
But there is still problems.

First of all, bands dyed with SS disappeared after 10sec.(312nm UV). Why? It's interesting that this is happening only in my lab. The friend of mine also use SS but it works just fine. Why is this happening only to me?

- the second, does anybody know why is SG so unusefull in my case? I have to tell, that I work with very old isolated DNA(4-5years). When I use fresh isolated DNA(under 1 year), it (SG) works fine. Why?
So, I found the suspect smile.gif
Thanks all for discussion.

-k4bf0389-

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