House dust mite extract from vendor AirMid
Bacterial 16S rRNA amplicons were constructed via amplification of the V4 region of the 16S rRNA gene with universal primers previously developed against the V4 region, flanked by Illumina standard adapter sequences.    Dual-indexed forward and reverse primers were used in all reactions.  Amplicon pools were combined, thoroughly mixed, and then purified.  The final amplicon pool was evaluated using the Advanced Analytical Fragment Analyzer automated electrophoresis system, quantified, and diluted according to Illuminas standard protocol for sequencing on the MiSeq instrument.