Posted 27 January 2009 - 06:54 AM
All the other samples were running from - to +. All samples were PCRed with the same chemicals at the same time by the same person
Anybody an idea what could have happened there?????? And why only 4 samples were going the wrong way????
Posted 27 January 2009 - 08:15 AM
is there anything that was different about how those samples were prepared (eg pH) that might have caused a charge reversal of the nucleic acid? or neutralization?
if you somehow neutralized the nucleic acid (eg by creating a salt) then the migration may have been by electroosmotic flow instead of by charge. eof generally runs from + to -.
if you used a low eof agarose then the sample may have remained at the origin.
genius does what it must
i do what i get paid to do
Posted 27 January 2009 - 08:56 AM
But the four samples really were running from + to - including both dyes from the loading buffer......and I tried again: same picture
But thanks for your suggestion, it sounds rational. Still puzzeld how something can happen!
Posted 27 January 2009 - 09:49 AM
Posted 27 January 2009 - 11:31 AM
Posted 27 January 2009 - 11:23 PM
no its a horizontal gel......and I do not think that its EtOH contamination, but I repeated the PCR and hope it works this time....and will repeat the extraction, maybe just some sort of my samples feeling the snow coming