16S-Seq of T-shirt:Human worn
Sequencing libraries were prepared using  a 15 cycle PCR targeting variable regions V3 and V4 of the 16S rRNA gene,  approx. 460bp, using primers (Uni341F: 5-CCTAYGGGRBGCASCAG-3 and Uni806R:  5-GGACTACNNGGGTATCTAAT-3) which also included sequencing adaptors and  barcode tags. PCR reactions were performed using PCRBIO HiFi polymerase,  modified to 25 l reactions (with 2 L nuclease free H2O), following the  manufacturers instructions. Amplicon products were purified using HighPrep  PCR (AC-60500, MagBio Genomics Inc., USA) PCR Clean Up System (0.65 L  beads:1 L amplicon product). Samples were normalized using SequalPrep  Normalization Plate (96) Kit (Invitrogen, Maryland, MD, USA), pooled, and  then concentrated using the DNA Clean and Concentrator-5 kit (Zymo Research,  Irvine, CA, USA). The pooled library concentration was determined using the  Qubit dsDNA HS Assay Kit (Invitrogen, Thermo Scientific) and adjusted to  1.65 ng/l (4 nM). Paired end Amplicon sequencing was performed on the  Illumina MiSeq Desktop Sequencer (Illumina Inc., CA, US) with the denatured  libraries adjusted to a final concentration of 16 pM. For each run, 5.0% PhiX  was included as internal control and the MiSeq Reagent Kits v2 were used  (Illumina Inc., CA, US). MiSeq Controller Software was used for sequence  demultiplexing.