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Sample Preparation | Purification of Genomic DNA

Meta-G-Nome™ DNA Isolation Kit

Order | Protocol

Applications

  • Isolation of fosmid cloning-ready metagenomic DNA from microbes in water, soil, and compost samples.
  • Isolation of metagenomic DNA for PCR or next-generation sequencing.

Note: The Meta-G-Nome™ DNA Isolation Kit replaces EPICENTRE's Metagenomic DNA Isolation Kit for Water.

The Meta-G-Nome™ DNA Isolation Kit is used to isolate inhibitor-free, fosmid cloning-ready DNA from unculturable or difficult-to-culture microbial species present in environmental water, soil, or compost samples (Fig. 1). The kit isolates high-molecular-weight DNA that is randomly sheared (with fragment sizes approximately 40 kb), does not require size selection, and can be used directly for end-repair and construction of fosmid libraries, PCR amplification, or next-generation sequencing. The kit can also be used to prepare 16S rRNA metagenomic libraries for phylogenetic classification or species-abundance studies.

The procedure uses filtration technology (with 1.2-µm and 0.45-µm membranes provided) and enzymatic lysis (with Ready-Lyse™ Lysozyme and Proteinase K) to isolate metagenomic DNA from water (100 ml per extraction), soil (1 g per extraction), or compost (300 mg per extraction). The purified DNA is free from humic and fulvic acids.

Benefits

  • No bead-beating, agarose plugs, pulsed-field gel electrophoresis, or size selection.
  • No phenol/chloroform extractions or CTAB.

Evaluation Program

  • Fill out a short online survey, and qualify for 50% off your first kit. Evaluation program is for U.S. customers only.
Figure 1

Figure 1 (click to enlarge). Schematic overview of the procedure for isolating DNA from soil or water samples using the Meta-G-Nome™ DNA Isolation Kit.
Figure 2 Figure 3
Figure 2. Size of inserts from a metagenomic fosmid library. Meta-genomic DNA isolated from lake water was used to generate a fosmid library using the CopyControl™ Fosmid Library Production Kit. DNA was isolated from randomly selected clones and digested with Not I. The insert sizes were approximately 40 kb. Lane M, Kilobase ladder; lane 1, 40-kb control insert; lanes 2-19, Not I-digested fosmid DNA. Figure 3. PCR amplification of soil metagenomic DNA. DNA was isolated following the kit protocol, and PCR was performed using 16S rRNA primers using various dilutions of the template DNA. Lane M, Kilobase ladder; lanes 1 and 2, 1 µl and 5 µl of PCR products from 1:10 template dilution; lanes 3 and 4, 1 µl and 5 µl of PCR products from 1:100 template dilution; lane 5, 1 µl of PCR products from undiluted template; lane 6, 1:10 dilution of sample from lane 5.

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   Catalog No. Size
Meta-G-Nome™ DNA Isolation Kit
   MGN0910 10 Purifications
Note: This kit requires a suitable filtration apparatus (e.g., MilliPore Cat. No. XX1004700, Nalgene Cat. No. 130-4045), which is not included.
Contains: Extraction buffer, Filter Wash Buffer, Meta-Lysis Solution (2X), MPC Protein Precipitation Reagent, Proteinase K, Fosmid Control DNA, RNase A, Ready-Lyse™ Lysozyme Solution, TE Buffer, 0.45-µm cellulose acetate filter membranes, 1.2-µm cellulose acetate filter membranes.

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