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DNA contamination? - PCR problems

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Old 10-18-2007, 02:52 PM
Pipette Filler
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Question DNA contamination? - PCR problems
Hi,
I have some problems with DNA extracted from frozen Papaver leaves.
During the DNA extraction (CTAB or Phenol protocols) my samples becames brown and after it's imposible to amplify by PCR, using RAPDs primers.
I think any product originated by oxidation prevents polimerase action.
Any idea of another DNA extration protocol? Or a way to purify/clean these samples of DNA? Help!!!
Susana
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Old 10-18-2007, 03:31 PM
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Default Re: DNA contamination? - PCR problems
Either Isoamyl alcohol:chloroform (24:1) after phenol extraction step as many times as needed to rid yourself of the brown color, then salt precipitation with NaAc, EtOH wash etc., or Chromaspin-100 column purification after any extraction protocol (repeat using two columns total).
Last edited by danfive; 10-18-2007 at 04:35 PM.
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Old 04-25-2008, 09:04 AM
Pipette Filler
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Default Re: DNA contamination? - PCR problems
have you tried edwards preps?

I havent had any problems amplifying dna from any plant tissue yet. *knocks on wood*
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