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    Zymo-Seq miRNA Library Kit

    R3006 / R3007

    Zymo-Seq miRNA Library Kit

    Cat # Name Size
    R3006 Zymo-Seq miRNA Library Kit 12 preps
    R3007 Zymo-Seq miRNA Library Kit 96 preps



    • Accurate miRNA profiling: A unique single adapter and circularization strategy reduces ligation bias and increases miRNA detection.
    • Direct detection: Generates small RNA libraries without the need for small RNA enrichment.
    • Gel-Free: Removes adapter dimers from any input without lengthy gel excision.

    The Zymo-Seq™ miRNA Library Kit is an innovative small RNA sequencing library prep kit that uses a single-adapter and circularization strategy to reduce ligation bias and provide accurate small RNA profiling from cell-free RNA (cfRNA) and total RNA. This simple, streamlined workflow minimizes hands-on time and uses gel-free removal of adapter dimers to generate stranded small RNA libraries from plasma, biofluids, cells, tissues, and more.

    To prepare Zymo-Seq™ miRNA libraries, RNA is ligated to the single miRNA adapter and the ligated product is circularized. Excess adapter is blocked and removed with specialized magnetic beads that allow a completely gel-free workflow for input as low as 1 ng. The circularized miRNA-adapter product is then reverse transcribed. The resulting cDNA is amplified using Indexing PCR primers which add on Illumina-compatible adapter and unique dual index (UDI) sequences for cost-effective multiplexing. Everything needed to make sequencing-ready libraries, including indexing primers and SPRI beads, are included in the kit. It’s RNA-Seq made simple.

    Additional Components Required (Not Provided) Molecular biology grade pure ethanol (> 95%)
    Maximum Input 200 ng of total RNA, or RNA extracted from 200 µL of plasma or serum.
    Minimum Input 1 ng of total RNA, or RNA extracted from 50 µL of plasma or serum.
    Processing Time 2 hours hands-on, 8 hours total
    Recommended Equipment A 96-well Aluminum Cooler Block for PCR tubes or a 96-well PCR tube rack may be placed on ice for ease of reagent assembly and maintaining samples at 4°C. To process > 12 samples at once, a multichannel pipette with low-retention tips is recommended for mixing and the timely addition of pre-aliquoted reagents.
    Recommended Input 100 ng of total RNA, or RNA extracted from 200 µL of plasma or serum.
    Required Equipment Thermal cycler, magnetic stand for 0.2 mL PCR tubes or 96-well plate, mini centrifuge, and micro centrifuge.
    Sample Type High integrity total RNA (RIN > 7) containing miRNA, or cell-free RNA extracted from plasma/serum samples not preserved with heparin. Small RNA enrichment is not necessary.
    Sequencing Platform Compatibility Libraries are compatible with all Illumina sequencing platforms.

    The Zymo-seq miRNA Library Kit can be used with a variety of RNA samples that contain microRNA, including plasma, urine, saliva, exosomes, and total RNA extracted from various cells and tissues. Many purification methods do not adequately purify small RNA. It is critical that the RNA purification method used effectively retains the microRNA and small RNAs, such as Direct-zol Microprep, Quick-RNA Microprep, and Quick-cf-RNA Serum & Plasma Kit.

    When using total RNA, we recommend using high quality RNA with a RNA Integrity Number (RIN) value of at least 7. RNA with a lower RIN value may be used but may result in over-representation of rRNA sequences due to the presence of higher levels of fragmented rRNA. This could reduce the representation of miRNAs and other small RNAs in the final library.

    The total RNA may be used without size selection. RNAs larger than ~100 nt within the sample are not effectively incorporated into the final library.

    Zymo-Seq miRNA libraries are compatible with any Illumina® sequencer, as well as the Element Biosciences AVITI™ system (with Element Adept Library Compatibility workflow.). This kit has not been validated on Ion Torrent®, Oxford Nanopore®, PacBio®, or other non-Illumina sequencing platforms.

    Libraries may be sequenced at any length. However, it is important to note that reads longer than the typical insert size (~21nt-100nt) will require more adapter trimming through the excess sequencing cycles. For optimal results, we recommend sequencing with a minimum read length of 50 base pairs (bp) and a maximum of 75 bp and single-end sequencing.

    We are happy to offer additional recommendations, troubleshooting assistance, and consultation requests. Email us at if you require further guidance or have any questions regarding the Zymo-seq miRNA Library Kit.

    "Easy handling, Less library prep time and less steps with Dimer removal bead"

    - Dr. Ramji Bhandari from the University of North Carolina of Greensboro

    "I am really happy with the kit! This experience with Zymo Kit was wonderful. Congratulations on the excellent product developed. I'm sure it will be a success. I had excellent library performance on the tests.”

    - Dr. Fabio Queiroz from the Instituto Mario Penna – MG/Brazil

    “Stress-free. Manageable. Fast. I really love the kit. The Zymo miRNA library kit dispelled previous discouraging experiences that I had when using individual components as described by different research articles. Every step is clearly explained in the manual and in the tutorial video, thus reading through the manual / watching the tutorial video prior to small RNA library preparation is highly recommended. It made the whole process simple and easy.”

    - Kim Nguyen, PhD, UC Irvine

    Cat # Name Size
    E2003 ZymoTaq PreMix (50 Rxns.) (2 x 625 µl)
    E2004 ZymoTaq PreMix (200 Rxns.) (8 x 625 µl)
    W1001-1 DNase/RNase-Free Water 1 ml
    W1001-4 DNase/RNase-Free Water 4 ml
    W1001-6 DNase/RNase-Free Water 6 ml
    W1001-10 DNase/RNase-Free Water 10 ml
    W1001-30 DNase/RNase-Free Water 30 ml
    D4003-2-6 DNA Wash Buffer (Concentrate) 6 ml
    D4003-2-48 DNA Wash Buffer (Concentrate) 48 ml
    D4003-2-24 DNA Wash Buffer (Concentrate) 24 ml
    D3004-4-10 DNA Elution Buffer 10 ml
    D3004-4-50 DNA Elution Buffer 50 ml
    3DP-1002 PCR Strip MagStand 1 unit