HIGHLIGHTS
- 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.
DESCRIPTION
The Quick-16S Plus NGS Library Prep Kit (V1-V2) is the fastest and simplest NGS library prep targeting the V1-V2 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.
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TECHNICAL SPECIFICATIONS
16s V1-V2 Primer Sequences (adapters not included) |
27f (AGRGTTYGATYMTGGCTCAG, 20 bp) and 341r (CTGCWGCCHCCCGTAGG, 17 bp). |
Amplicon Size |
The final amplicon size after 1-Step PCR (targeted amplification and barcode addition) is ~492 bp. |
Barcode Sequences |
10 bp barcodes are available to download here or by visiting the Documentation section of this Product Page. |
Index Primers |
Unique dual index (barcodes) to label individual samples. |
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® or NextSeq 1000/2000® without the need to add custom sequencing primers. We recommend the MiSeq® Reagent Kit v3 (600-cycle) or NextSeq 1000/2000® Reagent Kit P1/P2 (600-cycle). For assistance with sample sheet setup, see Appendix F. |