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Blotchy Blots


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#1 biochem06

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Posted 12 April 2016 - 02:25 AM

Hi

 

I'm wondering does anyone know what would be causing these "blotchy" blots. Semi dry transfer, all new reagents, new buffer etc. Transferred successfully as rainbow markers all transferred. You can see in the images when I ponceau stain there is this white patch in the middle. This is happening all the time. When I develop the blots there is always a high background/they are dirty.

 

Any ideas?

 

thanks

Attached Thumbnails

  • ponceau blot 2.jpg
  • ponceau stained blot.jpg


#2 mdfenko

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Posted 12 April 2016 - 04:06 AM

a few possible reasons:

 

poor contact in that area during transfer

 

shaking during staining/destaining is insufficient, allowing membrane to stick to staining vessel

 

pouring destain solution (water when using ponceau) directly onto blot at that spot


talent does what it can
genius does what it must
i do what i get paid to do

#3 biochem06

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Posted 12 April 2016 - 07:28 AM

many thanks mdfenko.






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