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Problematic DNA Extraction

extraction pcr dna

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#1 hmwmx

hmwmx

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Posted 14 April 2017 - 08:24 AM

Hello, 

 

Our lab has noticed some green/black contamination in a solution of 30% (w/v) PEG 400, 0.1M Na HEPES (pH 7.5), and 0.2M MgCl hexahydrate which we believe to be bacterial or fungal (or both) and we would like to attempt PCR sequencing in order to identify the offending microorganism.

 

Unfortunately, extracting any DNA (if present) is proving rather difficult. We have attempted mechanical lysis using glass beads, however, that has not yielded any positive results.

 

I think we'll likely spin the contamination down, re-suspend the resulting pellet in PBS and attempt enzymatic lysis, however, I thought I would enquire as to whether anyone have any experience of extracting bacterial/fungal DNA from unusual chemical solutions?

 

If anyone has any literature in mind some direction would be greatly appreciated...

 

Thanks.







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