- 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
The Quick-16S Plus NGS Library Prep Kit (V3-V4) 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.
Amplicon Size - The final amplicon size after 1-Step PCR (targeted amplification and barcode addition) is ~606 bp.
Barcode Sequences - 10 bp barcodes, Available for download here (USA Only), or under the Documents section as "Barcode Sequences".
Index Primers - Dual index (barcodes) to uniquely label 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.