Quick-16S Plus NGS Library Prep Kit
(V3-V4, UDI), w/Primer Set 3, Rxns 96
Zymo D6421-PS3

Quick-16S Plus NGS Library Prep Kit
(V3-V4, UDI), w/Primer Set 3, Rxns 96
Zymo D6421-PS3

The Quick-16S Plus NGS Library Prep Kit (V3-V4, UDI) is the fastest and simplest NGS library prep targeting the V3-V4 region of the 16S rRNA gene for high-throughput sequencing. The automation-friendly protocol utilizes a single qPCR/PCR for combined targeted amplification and barcode addition using specially designed primers. After pooling by equal volume, a single clean-up of the final library is performed, rather than massive AMPure® bead-based clean-ups. Additional library quantification analysis such as TapeStation analysis or gel electrophoresis are not necessary. With these features, the workflow dramatically reduces the hands-on time of library preparation to only 30 minutes.

The most streamlined NGS kit with only 30 minutes of hands-on time for 96 samples. 100% automation ready with only a single PCR step and without the need for normalization. Real-time PCR enables absolute microbial copy number quantification.

16S V3-V4 Primer Sequences (adapters not included): 341f (CCTACGGGDGGCWGCAG, CCTAYGGGGYGCWGCAG, 17 bp) and 806r (GACTACNVGGGTMTCTAATCC, 24 bp). The forward primer 341f is a mixture of the two sequences listed. Amplicon Size: The final amplicon size after 1-Step PCR (targeted amplification and barcode addition) is ~606 bp. Barcode Sequences: Barcode Sequences: 10 bp barcodes, Available for download here. Index Primers: Unique dual index (barcodes) to label individual samples. Primer Coverage: bacteria (84.2%), archaea (75.8%), and eukaryota (0.0%). Required Equipment: Microcentrifuge, plate spinner (centrifuge), 96-well real-time quantitative PCR system (SYBR Green compatible) or standard PCR system, and 96-well real-time PCR plates. Sample Input: Purified microbial DNA ≤100 ng, free of PCR inhibitors. Sequencing Platform: Illumina MiSeq without the need to add custom sequencing primers. Zymo Research recommends the MiSeq Reagent Kit v3 (600-cycle). For assistance with sample sheet setup, see Appendix F.