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  • Control Reaction (E2612)

    Included in the kit are control reagents and primers to detect human ribosomal protein gene, RPL30, and a 16s RNA universal region in E. coli DNA, as well as a sample of human IMR-90/E. coli (10:1 ratio) DNA. (Sequences of the primers and target DNA are listed on the manual pages 16–17). A typical control reaction that will yield a sufficient quantity of eluted material to perform an agarose gel assay or qPCR reactions at least in triplicate is outlined below:

    1. Perform capture experiment using 2.5 μl (250 ng) of IMR90/E. coli DNA as described on the manual pages 5–7.

    2. Carefully transfer the supernatant containing the target microbial DNA to a fresh microcentrifuge tube, and perform AMPure Bead Clean-up or Ethanol Precipitation. This is your enriched microbial DNA sample.

    3.  Wash bead pellet (Step 2 in "Collect Enriched Microbial DNA" Section) and elute the DNA by incubation at 75°C in 50 μl of TE plus Proteinase K, as described here .

    4. Carefully remove the supernatant to a fresh microcentrifuge tube, and perform AMPure Bead Clean-up or Ethanol Precipitation. This is your host DNA sample.