ITS seq of cement biofilm
For bacterial community analysis, the V3-V4 hypervariable region of the 16S rRNA gene was amplified using the primers 341F (5'-CCTACGGGNGGCWGCAG-3') and 806R (5'-GACTACHVGGGTATCTAATCC-3') [20][21]. For fungal community analysis, the internal transcribed spacer (ITS) region was amplified using the primers ITS1F (5'-CTTGGTCATTTAGAGGAAGTAA-3') and ITS2R (5'-GCTGCGTTCTTCATCGATGC-3') [22][23]. Amplicon sequencing was performed on an Illumina MiSeq platform (Illumina Inc., San Diego, CA, USA) with paired-end (2  300 bp) sequencing at the Genomics and Microbiome Core Facility at Rush University, USA.