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. |
Order information
| Cat # | Name | Size |
|---|---|---|
| D6421-PS1 | Quick-16S Plus NGS Library Prep Kit (V3-V4 UDI) with Primer Set 1 | 96 rxns |
| D6421-PS2 | Quick-16S Plus NGS Library Prep Kit (V3-V4, UDI) with Primer Set 2 | 96 rxns |
| D6421-PS3 | Quick-16S Plus NGS Library Prep Kit (V3-V4, UDI) with Primer Set 3 | 96 rxns |
| D6421-PS4 | Quick-16S Plus NGS Library Prep Kit (V3-V4, UDI) with Primer Set 4 | 96 rxns |
| D6421-PS5 | Quick-16S Plus NGS Library Prep Kit (V3-V4, UDI) with Primer Set 5 | 96 rxns |
| D6421-PS6 | Quick-16S Plus NGS Library Prep Kit (V3-V4, UDI) with Primer Set 6 | 96 rxns |
| D6421-PS7 | Quick-16S Plus NGS Library Prep Kit (V3-V4, UDI) with Primer Set 7 | 96 rxns |
| D6421-PS8 | Quick-16S Plus NGS Library Prep Kit (V3-V4, UDI) with Primer Set 8 | 96 rxns |
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