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Sample loading buffer turning yellow.


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#1 Aravin

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Posted 23 June 2010 - 01:48 PM

I added sample loading buffer (Bromophenol blue) to my sample and after boiling for 5 min, it turned yellow. Does anyone know the explanation behind this?

Thanks.

#2 Inmost sun

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Posted 24 June 2010 - 02:29 AM

I added sample loading buffer (Bromophenol blue) to my sample and after boiling for 5 min, it turned yellow. Does anyone know the explanation behind this?

Thanks.


Actually, BPB is a pH indicator turning yellow at acidic pH; so, your sample contains some protons which were not buffered by the sample buffer. Was it blue before boiling? Why so long boiling?

Edited by Inmost sun, 24 June 2010 - 02:31 AM.


#3 Prep!

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Posted 24 June 2010 - 08:42 PM

I added sample loading buffer (Bromophenol blue) to my sample and after boiling for 5 min, it turned yellow. Does anyone know the explanation behind this?

Thanks.


Actually, BPB is a pH indicator turning yellow at acidic pH; so, your sample contains some protons which were not buffered by the sample buffer. Was it blue before boiling? Why so long boiling?


i second tat.. ur sample had acidic pH!!!
once u load it and the gel starts running it will again turn blue cause of the pH of the lamelli buffer!!!

Edited by Prep!, 24 June 2010 - 08:42 PM.

Support bacteria - They are the only culture some people have!!!
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#4 zienpiggie

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Posted 29 June 2010 - 01:38 PM

Stumbled upon this when googling for my own stuff.

31. Sample preparation yellow in colour

a) Solution acidic: add NaOH until the solution turn blue.

:P Too little bromophenol blue in sample buffer.


http://aldrin.tripod.com/index-3.html




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