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Soil Metagenome extraction

metagenome soil PCR

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3 replies to this topic

#1 Eslam Samir

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Posted 22 August 2014 - 02:33 PM

Does any one work in soil metagenome ?

I found very low yeild with A260/A280 = 1.5 !!! highly contaminated DNA with humic acid or protein or both

 

any one has a solution ?



#2 bob1

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Posted 22 August 2014 - 03:06 PM

These sorts of problems are common with these types of extractions, and are compounded by the fact that soils often contain clays which sequester cations, thereby preventing efficient extraction of the DNA. YOu could try using a plant DNA extraction method (manual, not kit) using Polyvinyl pyrrolidone (PVP) or something similar.



#3 Eslam Samir

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Posted 22 August 2014 - 03:34 PM

These sorts of problems are common with these types of extractions, and are compounded by the fact that soils often contain clays which sequester cations, thereby preventing efficient extraction of the DNA. YOu could try using a plant DNA extraction method (manual, not kit) using Polyvinyl pyrrolidone (PVP) or something similar.

I'm trying to extract metagenome of procaryots (Bacterial genomes) is that makes any difference ?

how could I use PVP to purify my extract ?



#4 bob1

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Posted 25 August 2014 - 12:11 AM

Just look around for PVP plant DNA extraction protocols. PVP is usually used to remove polyphenolics that are common in plant extractions, so they should work to remove similar stuff in soil extractions.







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