Nanoliter-scale next-generation sequencing library mediated high-throughput 16s RRNA microbial community profiling: Supplementary Figures

Supplementary Figure 1. Nanoliter-scale next-generation sequencing library mediated high-throughput 16s RRNA microbial community profiling.

One-step amplification and next-generation sequencing library construction of the V3/V4 region of the 16S rRNA gene, using inner and outer pairs of primers. Forward primer 314F and reverse primer 806R of the inner primers were used for amplifying the V3/V4 region of the 16S rRNA gene. The outer primers containing spacer sequence and spacer primer (SP) sequence at the 5’ region of the primer were used to construct SNAP-TE libraries compatible for Illumina MiSeq/HiSeq platform. Spacer sequences are used to improve the sequencing quality and comprise differential sequences with different lengths of bases. The SP sequence is both the amplification primer region of the outer primer and the sequencing primer of the Illumina platform.

Article Abstract: An ultra-high-throughput workflow of next-generation sequencing library construction at nanoliter scale for amplicon sequencing, termed Smartchip nanowell platform for target enrichment (SNAP-TE), was established with the concerted usage of a nano-dispenser system and a nanoliter-scale PCR chip. To demonstrate its advantages of overall cost and time for library construction, quality control, and pooling for large-scale samples over the time-consuming conventional method, target amplicon sequencing of widely used 16S ribosomal RNA gene V3-V4 region for microbial community profiling was chosen for comparison. The result of finding no significant difference in microbial community profiling between these two methods strongly supported that SNAP-TE is a cost-effective method for next-generation sequencing library construction for large-scale samples to conduct amplicon sequencing based applications.