title: Negative DNA extraction control
design: DNA negative extraction controls were amplified using primers specific to the V4 variable region of the 16S rRNA gene and sequenced at the University of Connecticut Microbial Analysis, Resources, and Services (MARS) core lab using standard protocols. In brief, indexed 515F and 806R primers and GoTaq (Promega, Madison, WI, USA) with the addition of 10 mg BSA were used for PCR amplification. The PCR reaction was incubated at 95C for 2 minutes, the 30 cycles of 30 s at 95.0C, 60 s at 50.0 C, and 60 s at 72.0 C, followed by a final extension at 72.0 C for 10 minutes. PCR products were quantified and visualized using the QIAxcel DNA Fast Analysis (Qiagen, Hilden, Germany). PCR amplicons were sequenced on the Illumina MiSeq (Illumina, San Diego, CA, USA) platform at the University of Connecticut MARS core lab following library preparation.
identifiers.namespace: SUB15058271
identifiers.id: E01Plate2