The Quick-16S Plus NGS Library Prep Kit (V4) is the fastest and simplest NGS library prep targeting the V4 region of the 16S rRNA gene for high-throughput sequencing. The automation-friendly protocol utilizes a premixed amplification plate and a single qPCR/PCR for combined targeted amplification and barcode addition. Pooling by equal volume and a single clean-up of the final library, rather than massive AMPure® bead-based clean-ups, simplifies the library prep process. 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.
– Fast: 8x less hands-on time than conventional 16S library prep protocols.
– Easy: Premixed Plate minimizes hands-on time and handling errors.
– Simple Workflow: 100% Automation ready with only a single PCR step and no need for normalization. Just pool by equal volume.
| Amplicon Size | The final amplicon size after 1-Step PCR (targeted amplification and barcode addition) is ~388 bp. |
|---|---|
| Equipment Needed (user provided) | Microcentrifuge, plate spinner (centrifuge), 96-well real-time quantitative PCR system (SYBR Green compatible recommended), or standard PCR system, and 96-well real-time PCR plates. |
| Index Sequences | 10 bp indexes are listed in the MiSeq Sample Sheet Template provided. |
| Sample Input | Purified microbial DNA (≤100 ng), free of PCR inhibitors. |
| Sequencing Platform | Compatible with all Illumina® sequencing platforms using custom sequencing primers. We recommend the MiSeq® Reagent Kit v2 (300-cycle). |
| V4 Primer Sequences (adapters not included) | 515f (GTGYCAGCMGCCGCGGTAA) and 806r (GGACTACNVGGGTWTCTAAT). |
Order information
| Cat # | Name | Size |
|---|---|---|
| D6430-PS1 | Quick-16S Plus NGS Library Prep Kit (V4) with Primer Set 1 | 96 rxns |
| D6430-PS2 | Quick-16S Plus NGS Library Prep Kit (V4) with Primer Set 2 | 96 rxns |
| D6430-PS3 | Quick-16S Plus NGS Library Prep Kit (V4) with Primer Set 3 | 96 rxns |
| D6430-PS4 | Quick-16S Plus NGS Library Prep Kit (V4) with Primer Set 4 | 96 rxns |
| D6432 | Quick-16S Plus NGS Library Prep Kit (V4) | 384 rxns |
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