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    Quick-16S Plus NGS Library Prep Kit (V1-V2)


    Zymo Research's 100% Satisfaction Guarantee

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    Cat # Name Size
    D6434-PS1 Quick-16S Plus NGS Library Prep Kit (V1-V2) with Primer Set 1 96 rxns

    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 & Documents Specifications FAQ Components Related Products

    Documents

    Product 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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    Documents

    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.

    Resources

    Purified total DNA from any organism free from PCR inhibitors. This system has been used by Zymo Research’s Microbiomics Services team to process samples from human, water, soil, food, plants, feces, biofilms, and much more.

    The system has been tested with inputs as low as 100 femtograms with no significant impact to the performance of the kit.

    This single PCR step combines targeted amplification of the microbial genome and the barcoding index PCR, which allows for a simple and fast workflow.

    The combination of using Equalase qPCR Premix to yield roughly equal amounts of libraries and a carefully curated barcode index list results in similar raw sequencing reads across samples.

    The ZymoBIOMICS™ Microbial Community DNA Standard consists of purified DNA from 8 bacteria and 2 yeasts and is useful for assessing community profile bias. The ZymoBIOMICS™ 16S/ITS qPCR Standard consists of a plasmid that has a single bacterial and single fungal target and is useful for quantifying copy number. Both are useful as positive controls for the library prep process.

    Equalase amplification and the library prep design may cause some library products to not anneal well, causing a lack of a tight band. These libraries are perfectly fine because preparation for Illumina sequencing includes denaturing libraries into single strands.

    Purified total DNA from any organism free from PCR inhibitors. This system has been used by Zymo Research’s Microbiomics Services team to process samples from human, water, soil, food, plants, feces, biofilms, and much more.

    The system has been tested with inputs as low as 100 femtograms with no significant impact to the performance of the kit.

    This single PCR step combines targeted amplification of the microbial genome and the barcoding index PCR, which allows for a simple and fast workflow.

    The combination of using Equalase qPCR Premix to yield roughly equal amounts of libraries and a carefully curated barcode index list results in similar raw sequencing reads across samples.

    The ZymoBIOMICS™ Microbial Community DNA Standard consists of purified DNA from 8 bacteria and 2 yeasts and is useful for assessing community profile bias. The ZymoBIOMICS™ 16S/ITS qPCR Standard consists of a plasmid that has a single bacterial and single fungal target and is useful for quantifying copy number. Both are useful as positive controls for the library prep process.

    Equalase amplification and the library prep design may cause some library products to not anneal well, causing a lack of a tight band. These libraries are perfectly fine because preparation for Illumina sequencing includes denaturing libraries into single strands.

    Cat # Name Size
    D4302-5-10 ZymoBIOMICS DNase/RNase Free Water 10 ml
    D6305 ZymoBIOMICS Microbial Community DNA Standard 200ng
    D6306 ZymoBIOMICS Microbial Community DNA Standard 2000ng

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