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  • Low DNA concentration from environmental samples

    Hi,

    I have an environmental sample that contains a mixed microbial population. The sample has been subjected to very harsh conditions, where after I isolated DNA. The idea is to do whole metagenome sequencing on both the environmental sample and after the harsh conditions to compare which communities survived. The problem I have is the DNA concentration is so low after the harsh conditions. I tried the Qubit (with 20 uL), but the concentration was too low to detect. I did PCR on the DNA and I obtained good amplicons for all three domains of life, so I know there is DNA, but not enough for whole metagenome sequencing. What can I do to improve this, without redoing the experiment. (It takes two months to repeat the experiment, and I don't have that time). I was looking at this WGA kit https://www.neb.com/products/e2620-picoplex-wga-kit but this is for single cell. I need something like this for a mixed community from environmental samples. Anyone have ideas please?

  • #2
    google searching "whole genome amplification kit" will find most of the commercially available amplification kits. Genome amplification will not discriminate between dead and live microorganism DNA and most of them are biased. So, after sequencing you may find it difficult to interpret the results.

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