5 replies to this topic

[siddharthsameer]

I forgot to add ethidium bromide in the molten agrose however i added the ethidium bromide to the running buffer...Will i be able to visualise the band, if not how caqn I rescue it. kindly let me know..
with regards

[bob1]

yes, it should be fine.

[siddharthsameer]

bob1, on 06 September 2012 - 12:19 AM, said:

yes, it should be fine.

thanks i added 5µl of ethidium bromide to the running buffer but i am tensed wheether i will be able to see it since i have to cut the band for the ligation...are you sure it will be fine.. thanks for your reply..

[metionina]

I used to visualize my bands after migration with an immersion of the gel in a solution with a high concentration of ethidium bromide for about 30 minutes. That works fine! In the running gel, probably would be fine but I think it depends also on the concentration of your sample. So, what is the result?

[siddharthsameer]

metionina, on 06 September 2012 - 02:20 AM, said:

I used to visualize my bands after migration with an immersion of the gel in a solution with a high concentration of ethidium bromide for about 30 minutes. That works fine! In the running gel, probably would be fine but I think it depends also on the concentration of your sample. So, what is the result?

It worked perfectly fine

[bob1]

It should do... lots and lots of people do this around the world.  Actually, all you need to do is add the EtBr to the +ve electrode end, it migrates in the opposite direction to DNA so will be drawn into the gel when running.