In the laboratory, the meconium was divided into 5 aliquots (each 200-mg) and immediately stored at −80 °C. A frozen aliquot (200 mg) of each fecal sample was processed using the QIAamp DNA Stool Mini Kit (QIAGEN, Hilden, Germany) for DNA extraction as previously described43 (link). The DNA concentration was measured with a NanoDrop (Thermo Scientific), and the molecular size was estimated by agarose gel electrophoresis.
Metagenomic DNA libraries were constructed with 0.2 μg of genomic DNA according to the Illumina TruSeq DNA Sample Prep v2 Guide, with an average insert size of 500 bp. The quality of all libraries was evaluated using an Agilent Bioanalyzer with a DNA LabChip 1000 kit. Negative controls (sterile water) were included for all the experimental process and showed no amplification in the final DNA library. Sequencing was performed using an Illumina Hiseq2500.
Metagenomic DNA libraries were constructed with 0.2 μg of genomic DNA according to the Illumina TruSeq DNA Sample Prep v2 Guide, with an average insert size of 500 bp. The quality of all libraries was evaluated using an Agilent Bioanalyzer with a DNA LabChip 1000 kit. Negative controls (sterile water) were included for all the experimental process and showed no amplification in the final DNA library. Sequencing was performed using an Illumina Hiseq2500.
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