16S rRNA gene sequencing of NICU environmental swab: rm_1_Drain
Libraries for sequencing were prepared according to Illumina s  16S Metagenomic Sequencing Library Preparation  guide, with the exception of using Qiagen HotStar Taq DNA polymerase for the first PCR ( amplicon PCR ). The template specific primers were (without the overhang adapter sequence): 343F (5 -TACGGRAGGCAGCAG-3 ) and 803R (5 -CTACCAGGGTATCTAATCC-3 ). The first PCR ( amplicon PCR ) was carried out for 32 cycles with annealing temperature of 55 C. Diluted pooled samples were loaded on an Illumina MiSeq and sequenced using a 600-cycles MiSeq Reagent Kit v3.