HIGHLIGHTS
- 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!
DESCRIPTION
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.
TECHNICAL SPECIFICATIONS
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). |
FORMAT
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 |