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PCR contamination


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19 replies to this topic

#16 tantao

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Posted 30 August 2010 - 06:02 AM

55C is not too high, although not too low either. If possible try a gradinent PCR, or if you cannot, try 60 C with a real sample and the contaminated water....

edit: try to clean the pipett in the inside as well!

Dear gebirgsziege,
how to clean the pipett in the inside. I use 75% alcohol to clean the pipett. Is it ok? Thanks again!

#17 gebirgsziege

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Posted 30 August 2010 - 06:45 AM

your lab manager should have a manual for this; otherwise contact the manufacturers homepage how to open and clean the pipett. Then use DNA off or diluted bleach to rise the parts that can be rinsed (see manual), let them dry overnight and re-assemble the pipett.
A man cannot be too careful in the choice of his enemies. (Oscar Wilde)

#18 Mullet

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Posted 10 September 2010 - 03:42 AM

If you are using filter tips for everything, then you shouldnt need to open the pipette (although its always nice to have clean gear!).

What did you prepare the new primer stock in? If you can buy some sterilised water and use that for making you stocks (and working soln), it might help.

What state is your autoclave in? Maybe you tips are coming out dirtier than when they went in? Try a new box of tips...

just my 0.02

#19 Maddie

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Posted 15 September 2010 - 06:38 AM

Are you sure your primers can't amplify human DNA? have you tried to sequence your band?
Theory is when we know everything and nothing is working. Practice is when everything is working and nobody knows why. Here, we combine theory and practice. Nothing is working and nobody knows why.

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#20 nightingale

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Posted 15 September 2010 - 11:55 AM

first : hope u have solved this ...
second : have you tried cleaning the area you are working in, the PCR rack , .... etc with bleach 10 % ?

wishing you all the best

" The more you learn, the more you realize how little you know ... "




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